Modulators of mas-related g-protein receptor x2 and related products and methods

ABSTRACT

Methods are provided for modulating MRGPRX2 generally, or for treating a MRGPRX2 dependent condition more specifically, by contacting the MRGPRX2 or administering to a subject in need thereof, respectively, an effective amount of a compound having structure (I): 
     
       
         
         
             
             
         
       
     
     or a pharmaceutically acceptable salt, isomer, hydrate, solvate or isotope thereof, wherein D, W, Z, R 1 , R 2  and R 3  are as defined herein. Pharmaceutical compositions containing such compounds, as well as the compounds themselves, are also provided.

BACKGROUND Technical Field

The invention relates to modulators of the Mas-related G-protein coupledreceptor X2, to products containing the same, as well as to methods oftheir use and preparation.

Description of the Related Art

Mas-related G-protein receptors (MRGPRs) are a group of orphan receptorswith limited expression in very specialized tissues. Very little isknown about the function of most of these receptors. There are eightrelated receptors in this class expressed in humans, only four of whichhave readily identifiable orthologs in other species (i.e., MRGPR D, E,F and G). The other four receptors (MRGPR X1, X2, X3 and X4) havecounterparts in higher species including monkeys, dogs and horses, butthey do not have a single corresponding ortholog in rodents.

BRIEF SUMMARY

This invention is based, in part, on the identification that MRGPRX2 orMRGPRX2 ortholog modulator compounds. MRGPRX2 corresponds functionallyto mouse MRGPRB2 and dog MRGPRX2 in mast cells. MRGPRX2 and its orthologreceptors mediate disorders including pseudo-allergic reactionsincluding pseudo-allergic drug reactions, chronic itch (e.g., pruritus),inflammation disorders, pain disorders, skin disorders, wound healing,cardiovascular disease, and lung inflammation/COPD. In one embodiment,both MRGPRB2 and MRGPRX2 expression is largely restricted to mast cells.Mast cells are innate immune cells that primarily reside at sitesexposed to the external environment, such as the skin,oral/gastrointestinal mucosa and respiratory tract. Mast cells expressnumerous receptors that respond to mechanical and chemical stimuli. Uponactivation, classically by IgE, mast cells release pre-formed mediatorsfrom granules (e.g., histamine, proteases, and heparin) and newlysynthesized mediators (e.g., thromboxane, prostaglandin D2, leukotrieneC4, tumor necrosis factor alpha, eosinol chemotactor factor, andplatelet-activating factor) that elicit allergic and inflammatoryresponses. Histamine dilates post-capillary venules, activates theendothelium, and increases blood vessel permeability. This causes localedema, warmth, redness, and chemotaxis of other inflammatory cells tothe site of release. Histamine also contributes to neuronalsensitization that leads to pain or itch. MRGPRB2 and MRGPRX2 mediateimmunoglobulin E (IgE) independent activation of mast cells. MRGPRB2 andMRGPRX2 are receptors for (or sensitive to activation by) variousligands, including basic secretagogues (small cationic molecules),certain drugs (e.g., cationic peptidergic drugs), neuropeptides, andantimicrobial peptides, and thus are important for non-IgE mediatedpseudo-allergic reactions, inflammation, pain, and itch conditions. Mastcells may also contribute to the progression of autoimmune disorders bypromoting chronic inflammation in the local tissue microenvironment andultimately polarizing toward a T_(h)17 immune response. Thus, modulatingMRGPRX2 or MRGPRX2 ortholog allows for treatment of autoimmune diseases,pseudo-allergic drug reactions, pain, itch, and inflammatory disorderssuch as inflammatory bowel disease, urticaria, sinusitis, asthma,rosacea, endometriosis, and other MRGPRX2 or MRGPRX2 ortholog dependentconditions as explained in more detail below.

In one embodiment, a method is provided for treating a MRGPRX2 or aMRGPRX2 ortholog dependent condition by administering to a subject inneed thereof an effective amount of the pharmaceutical composition ofthe modulator compounds of the present invention.

Accordingly, in an embodiment, is provided a compound having structure(I):

or a pharmaceutically acceptable salt, isomer, hydrate, solvate orisotope thereof, wherein D, W, Z, R¹, R² and R³ are as defined herein.

In other embodiments, compounds are provided having any of thestructures (Ia), (Ia′), (Ib), (Ic), (Id), (Ie), (If) or (Ig) as definedherein, or a pharmaceutically acceptable salt, isomer, hydrate, solvateor isotope thereof.

In yet other embodiments, pharmaceutical compositions are providedcomprising a carrier or excipient and a compound having structure (I),or a pharmaceutically acceptable salt, isomer, hydrate, solvate orisotope thereof.

In one embodiment, pharmaceutical compositions are provided comprisingsubstructures of structure (I) with structures (Ia), (Ia′), (Ib), (Ic),(Id), (Ie), (If) or (Ig) as defined herein or a pharmaceuticallyacceptable salt, isomer, hydrate, solvate or isotope thereof.

In another embodiment, methods are provided for treating an MRGPRX2 orMRGPRX2 ortholog dependent condition by administering to a subject inneed thereof an effective amount of a compound having structure (I), ora pharmaceutically acceptable salt, isomer, hydrate, solvate or isotopethereof.

In one embodiment, the MRGPRX2 or MRGPRX2 ortholog dependent conditionis one or more of a pseudo-allergic reaction, itch associated condition,a pain associated condition, an inflammation-associated condition, or anautoimmune disorder.

In one embodiment, the methods of treating the MRGPRX2 or MRGPRX2ortholog dependent condition are provided which comprise administeringan effective amount of a compound of structure (I) or any of thestructures (Ia), (Ia′), (Ib), (Ic), (Id), (Ie), (If) or (Ig) as definedherein or a pharmaceutically acceptable salt, isomer, hydrate, solvateor isotope thereof.

In another embodiment, compounds are provided having one or more of thestructures disclosed herein, or a pharmaceutically acceptable salt,isomer, hydrate, solvate or isotope thereof.

DETAILED DESCRIPTION

As mentioned above, the invention relates to modulators of the MRGPRX2,to products containing the same, as well as to methods of their use andpreparation. This invention is based, in part, on the identification ofMRGPRX2 modulator compounds. Both MRGPRB2 and MRGPRX2 are expressed inmast cells and dorsal root ganglia. Both MRGPRX2 and MRGPRB2 arereceptors for (or sensitive to activation by) a diverse group ofligands, including basic secretagogues, certain drugs, neuropeptides,antimicrobial peptides, and thus are important for pseudo-allergicreactions, itch, pain, or inflammatory disorders upon exposure.

MRGPRs appear to be sensory receptors that recognize their externalenvironment to exogenous or endogenous signals/chemicals. Thesereceptors likely respond to multiple chemical ligands/agonists. Forexample, MRGPRX2 recognizes Compound 48/80, Substance P, mastoparan,icatibant, ciprofloxacin, and tracurium, as agonist signals. In certainembodiments, molecules of this invention modulate MRGPRX2 by functioningas inverse agonists that are capable of blocking multiple chemicalentities, and/or as competitive antagonists that can specifically blockindividual ligands. In one embodiment, such modulations are selectiveagainst other MRGPRs, such as MRGPRX1, X3 and/or X4.

Definitions

As used herein, the following terms have the meaning defined below,unless the context indicates otherwise.

“Modulating” MRGPRX2 means that the compound interacts with the MRGPRX2in a manner such that it functions as an inverse agonist to thereceptor, and/or as a competitive antagonist to the receptor. In oneembodiment, such modulation is partially or fully selective againstother MRGPRs, such as MRGPRX1, X3 and/or X4.

“MRGPR” refers to one or more of the Mas-related G protein coupledreceptors, which are a group of orphan receptors with limited expressionin very specialized tissues (e.g., in mast cells and dorsal rootganglia) and barrier tissues. There are eight related receptors in thisclass expressed in humans, only 4 of which have readily identifiableorthologs in other species (i.e., MRGPR D, E, F and G). The other fourreceptors (MRGPR X1, X2, X3 and X4) have counterparts in higher speciesincluding monkeys, dogs and horses, but they do not have a singlecorresponding ortholog in rodents.

“MRGPRX2,” also referred to as “MRGX2,” or “MGRG3,” refers to a memberof the MRGPR family that is expressed on mast cells and capable ofmediating IgE independent activation (e.g., mast cell degranulation) inresponse to ligand binding. An exemplary human MRGPRX2 amino acidsequence is set forth in Uniprot Q96LB1.

“MRGPRX2 ortholog” refers to genes in non-human species including dogsand horses that have evolved through speciation events. Orthologs ofhuman MRGPRX2 are determined by expression pattern and pharmacology.Studies of non-human species MRGPRX2 show gene clusters containing somehuman MRGPRX2 members, some with coding genes, and some showing sequenceidentity to human MRGPRX2.

“Effective amount” refers to a quantity of a specified agent sufficientto achieve a desired effect in a subject being treated with that agent.Ideally, an effective amount of an agent is an amount sufficient toinhibit or treat the disease without causing substantial toxicity in thesubject. The effective amount of an agent will be dependent on thesubject being treated, the severity of the affliction, and the manner ofadministration of the pharmaceutical composition. Methods of determiningan effective amount of the disclosed compound sufficient to achieve adesired effect in a subject will be understood by those of skill in theart in light of this disclosure.

“Alkyl” means a saturated or unsaturated straight chain or branchedalkyl group having from 1 to 8 carbon atoms, in some embodiments from 1to 6 carbon atoms, in some embodiments from 1 to 4 carbon atoms, and insome embodiments from 1 to 3 carbon atoms. Examples of saturatedstraight chain alkyl groups include, but are not limited to, methyl,ethyl, n-propyl, n-butyl, n-pentyl-, n-hexyl, n-heptyl, and n-octylgroups. Examples of branched alkyl groups include, but are not limitedto, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and2,2-dimethylpropyl groups. An unsaturated alkyl includes alkenyl andalkynyl as defined below.

“Alkenyl” means a straight chain or branched alkenyl group having from 2to 8 carbon atoms, in some embodiments from 2 to 6 carbon atoms, in someembodiments from 2 to 4 carbon atoms, and in some embodiments from 2 to3 carbon atoms. Alkenyl groups are unsaturated hydrocarbons that containat least one carbon-carbon double bond. Examples of lower alkenyl groupsinclude, but are not limited to, vinyl, propenyl, butenyl, pentenyl, andhexenyl.

“Alkynyl” means a straight chain or branched alkynyl group having from 2to 8 carbon atoms, in some embodiments from 2 to 6 carbon atoms, in someembodiments from 2 to 4 carbon atoms, and in some embodiments from 2 to3 carbon atoms. Alkynyl groups are unsaturated hydrocarbons that containat least one carbon-carbon triple bond. Examples of alkynyl groupsinclude, but are not limited to, ethynyl, propynyl, butynyl, pentynyl,and hexynyl.

“Halo” or “halogen” refers to fluorine, chlorine, bromine, and iodine.

“Hydroxy” refers to —OH.

“Cyano” refers to —CN.

Amino refers to —NH₂, —NHalkyl or N(alkyl)₂, wherein alkyl is as definedabove. Examples of amino include, but are not limited to —NH₂, —NHCH₃,—N(CH₃)₂, and the like.

“Haloalkyl” refers to alkyl as defined above with one or more hydrogenatoms replaced with halogen. Examples of lower haloalkyl groups include,but are not limited to, —CF₃, —CHF₂, and the like.

“Alkoxy” refers to alkyl as defined above joined by way of an oxygenatom (i.e., —O-alkyl). Examples of alkoxy groups include, but are notlimited to, methoxy, ethoxy, n-propoxy, n-butoxy, isopropoxy,sec-butoxy, tert-butoxy, and the like.

“Haloalkoxy” refers to haloalkyl as defined above joined by way of anoxygen atom (i.e., —O-haloalkyl). Examples of lower haloalkoxy groupsinclude, but are not limited to, —OCF₃, and the like.

“Cycloalkyl” refers to alkyl groups forming a ring structure, which canbe substituted or unsubstituted, wherein the ring is either completelysaturated, partially unsaturated, or fully unsaturated, wherein if thereis unsaturation, the conjugation of the pi-electrons in the ring do notgive rise to aromaticity. Examples of cycloalkyl include, but are notlimited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, and cyclooctyl groups. In some embodiments, the cycloalkylgroup has 3 to 8 ring members, whereas in other embodiments the numberof ring carbon atoms range from 3 to 5, 3 to 6, or 3 to 7. Cycloalkylgroups further include polycyclic cycloalkyl groups such as, but notlimited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, andcarenyl groups, and fused rings such as, but not limited to, decalinyl,and the like.

“Aryl” groups are cyclic aromatic hydrocarbons that do not containheteroatoms. Representative aryl groups include, but are not limited to,phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl,phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl,biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments,aryl groups contain 6-14 carbons in the ring portions of the groups. Theterms “aryl” and “aryl groups” include fused rings wherein at least onering, but not necessarily all rings, are aromatic, such as fusedaromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, andthe like). In one embodiment, aryl is phenyl or naphthyl, and in anotherembodiment aryl is phenyl.

“Carbocycle” refers to alkyl groups forming a ring structure, which canbe substituted or unsubstituted, wherein the ring is either completelysaturated, partially unsaturated, or fully unsaturated, wherein if thereis unsaturation, the conjugation of the pi-electrons in the ring maygive rise to aromaticity. In one embodiment, carbocycle includescycloalkyl as defined above. In another embodiment, carbocycle includesaryl as defined above.

“Heterocycle” refers to aromatic and non-aromatic ring moietiescontaining 3 or more ring members, of which one or more is a heteroatomsuch as, but not limited to, N, O, S, or P. In some embodiments,heterocyclyl include 3 to 20 ring members, whereas other such groupshave 3 to 15 ring members. At least one ring contains a heteroatom, butevery ring in a polycyclic system need not contain a heteroatom. Forexample, a dioxolanyl ring and a benzdioxolanyl ring system(methylenedioxyphenyl ring system) are both heterocyclyl groups withinthe meaning herein.

Heterocyclyl groups also include fused ring species including thosehaving fused aromatic and non-aromatic groups. A heterocyclyl group alsoincludes polycyclic ring systems containing a heteroatom such as, butnot limited to, quinuclidyl, and also includes heterocyclyl groups thathave substituents, including but not limited to alkyl, halo, amino,hydroxy, cyano, carboxy, nitro, thio, or alkoxy groups, bonded to one ofthe ring members. A heterocyclyl group as defined herein can be aheteroaryl group or a partially or completely saturated cyclic groupincluding at least one ring heteroatom. Heterocyclyl groups include, butare not limited to, pyrrolidinyl, furanyl, tetrahydrofuranyl,dioxolanyl, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, pyrazolyl,triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, thiophenyl, benzothiophenyl, benzofuranyl, dihydrobenzofuranyl, indolyl,dihydroindolyl, azaindolyl, indazolyl, benzimidazolyl,azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl,imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl,xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl,tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups.

In one embodiment, heterocyclyl includes heteroaryl.

“Heteroaryl” refers to aromatic ring moieties containing 5 or more ringmembers, of which, one or more is a heteroatom such as, but not limitedto, N, O, and S. Heteroaryl groups include, but are not limited to,groups such as pyrrolyl, pyrazolyl, pyridinyl, pyridazinyl, pyrimidyl,pyrazyl, pyrazinyl, pyrimidinyl, thienyl, triazolyl, tetrazolyl,triazinyl, thiazolyl, thiophenyl, oxazolyl, isoxazolyl, benzothiophenyl,benzofuranyl, indolyl, azaindolyl, indazolyl, benzimidazolyl,azabenzimidazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl,imidazopyridinyl, isoxazolopyridinyl, thianaphthalenyl, purinyl,xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl,tetrahydroquinolinyl, tetrahydroisoquinolinyl, quinoxalinyl, andquinazolinyl groups. The terms “heteroaryl” and “heteroaryl groups”include fused ring compounds such as wherein at least one ring, but notnecessarily all rings, are aromatic, including tetrahydroquinolinyl,tetrahydroisoquinolinyl, indolyl, and 2,3-dihydro indolyl.

“Isomer” is used herein to encompass all chiral, diastereomeric orracemic forms of a structure (also referred to as a stereoisomer, asopposed to a structural or positional isomer), unless a particularstereochemistry or isomeric form is specifically indicated. Suchcompounds can be enriched or resolved optical isomers at any or allasymmetric atoms as are apparent from the depictions, at any degree ofenrichment. Both racemic and diastereomeric mixtures, as well as theindividual optical isomers can be synthesized so as to be substantiallyfree of their enantiomeric or diastereomeric partners, and these are allwithin the scope of certain embodiments of the invention. The isomersresulting from the presence of a chiral center comprise a pair ofnonsuperimposable-isomers that are called “enantiomers.” Singleenantiomers of a pure compound are optically active (i.e., they arecapable of rotating the plane of plane polarized light and designated Ror S).

“Isolated optical isomer” means a compound which has been substantiallypurified from the corresponding optical isomer(s) of the same formula.For example, the isolated isomer may be at least about 80%, at least 80%or at least 85% pure by weight. In other embodiments, the isolatedisomer is at least 90% pure or at least 98% pure, or at least 99% pureby weight.

“Substantially enantiomerically or diastereomerically” pure means alevel of enantiomeric or diastereomeric enrichment of one enantiomerwith respect to the other enantiomer or diastereomer of at least about80%, and more specifically in excess of 80%, 85%, 90%, 95%, 98%, 99%,99.5% or 99.9%.

The terms “racemate” and “racemic mixture” refer to an equal mixture oftwo enantiomers. A racemate is labeled “(±)” because it is not opticallyactive (i.e., will not rotate plane-polarized light in either directionsince its constituent enantiomers cancel each other out). All compoundswith an asterisk (*) adjacent to a tertiary or quaternary carbon areoptically active isomers, which may be purified from the respectiveracemate and/or synthesized by appropriate chiral synthesis.

A “hydrate” is a compound that exists in combination with watermolecules. The combination can include water in stoichiometricquantities, such as a monohydrate or a dihydrate, or can include waterin random amounts. As the term is used herein a “hydrate” refers to asolid form; that is, a compound in a water solution, while it may behydrated, is not a hydrate as the term is used herein.

A “solvate” is similar to a hydrate except that a solvent other thatwater is present. For example, methanol or ethanol can form an“alcoholate”, which can again be stoichiometric or non-stoichiometric.As the term is used herein a “solvate” refers to a solid form; that is,a compound in a solvent solution, while it may be solvated, is not asolvate as the term is used herein.

“Isotope” refers to atoms with the same number of protons but adifferent number of neutrons, and an isotope of a compound of structure(I) includes any such compound wherein one or more atoms are replaced byan isotope of that atom. For example, carbon 12, the most common form ofcarbon, has six protons and six neutrons, whereas carbon 13 has sixprotons and seven neutrons, and carbon 14 has six protons and eightneutrons. Hydrogen has two stable isotopes, deuterium (one proton andone neutron) and tritium (one proton and two neutrons). While fluorinehas a number of isotopes, fluorine-19 is longest-lived. Thus, an isotopeof a compound having the structure of structure (I) includes, but notlimited to, compounds of structure (I) wherein one or more carbon 12atoms are replaced by carbon-13 and/or carbon-14 atoms, wherein one ormore hydrogen atoms are replaced with deuterium and/or tritium, and/orwherein one or more fluorine atoms are replaced by fluorine-19.

“Salt” generally refers to an organic compound, such as a carboxylicacid or an amine, in ionic form, in combination with a counter ion. Forexample, salts formed between acids in their anionic form and cationsare referred to as “acid addition salts”. Conversely, salts formedbetween bases in the cationic form and anions are referred to as “baseaddition salts.”

The term “pharmaceutically acceptable” refers an agent that has beenapproved for human consumption and is generally non-toxic. For example,the term “pharmaceutically acceptable salt” refers to nontoxic inorganicor organic acid and/or base addition salts (see, e.g., Lit et al., SaltSelection for Basic Drugs, Int. J. Pharm., 33, 201-217, 1986)(incorporated by reference herein).

Pharmaceutically acceptable base addition salts of compounds of theinvention include, for example, metallic salts including alkali metal,alkaline earth metal, and transition metal salts such as, for example,calcium, magnesium, potassium, sodium, and zinc salts. Pharmaceuticallyacceptable base addition salts also include organic salts made frombasic amines such as, for example, N,N′dibenzylethylenediamine,chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine(N-methylglucamine), and procaine.

Pharmaceutically acceptable acid addition salts may be prepared from aninorganic acid or from an organic acid. Examples of inorganic acidsinclude hydrochloric, hydrobromic, hydriodic, nitric, carbonic,sulfuric, and phosphoric acids. Appropriate organic acids may beselected from aliphatic, cycloaliphatic, aromatic, aromatic aliphatic,heterocyclic, carboxylic, and sulfonic classes of organic acids,examples of which include formic, acetic, propionic, succinic, glycolic,gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic,fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic,4-hydroxybenzoic, phenylacetic, mandelic, hippuric, malonic, oxalic,embonic (pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic,panthothenic, trifluoromethanesulfonic, 2-hydroxyethanesulfonic,p-toluenesulfonic, sulfanilic, cyclohexylaminosulfonic, stearic,alginic, βhydroxybutyric, salicylic, -galactaric, and galacturonic acid.

The compounds of the disclosure (i.e., compounds of structure (I) andembodiments thereof), or their pharmaceutically acceptable salts maycontain one or more centers of geometric asymmetry and may thus giverise to enantiomers, diastereomers, and other stereoisomeric forms thatare defined, in terms of absolute stereochemistry, as (R)- or (S)- or,as (D)- or (L)- for amino acids. Embodiments thus include all suchpossible isomers, as well as their racemic and optically pure forms.Optically active (+) and (−), (R)- and (S)-, or (D)- and (L)-isomers maybe prepared using chiral synthons or chiral reagents, or resolved usingconventional techniques, for example, chromatography and fractionalcrystallization. Conventional techniques for the preparation/isolationof individual enantiomers include chiral synthesis from a suitableoptically pure precursor or resolution of the racemate (or the racemateof a salt or derivative) using, for example, chiral high pressure liquidchromatography (HPLC). When the compounds described herein containolefinic double bonds or other centers of geometric asymmetry, andunless specified otherwise, it is intended that the compounds includeboth E and Z geometric isomers. Likewise, all tautomeric forms are alsoincluded.

Although pharmaceutically unacceptable salts are not generally useful asmedicaments, such salts may be useful, for example as intermediates inthe synthesis of compounds having the structure of Formula I, forexample in their purification by recrystallization.

As used herein, the phrase “MRGPRX2 dependent condition” means acondition where the activation, over sensitization, or desensitizationof MRGPRX2 by a natural or synthetic ligand initiates, mediates,sustains, or augments a pathological condition. For example, it is knownthat some cationic peptidergic drugs cause pseudo-allergic reactions inpatients MRGPRX2 is sensitive to (or activated by) secretagogues,cationic peptidergic drugs, including icatibant, leuprolide, organirelix, neutral and anionic peptidergic drugs (e.g., exenatide,glucagon, liraglutide, enfuviritide, colistimethate), non-steroidalagonist (atracurium mivacurium), non-steroidal antagonist drugs,neuropeptides, and antimicrobial peptides. Moreover, overexpression ofMRGPRX2 and/or overactivity MRGPRX2 may also render mast cells moresusceptible to activation by endogenous and/or exogenous ligands.Without limited by theory, it is to be understood that by modulatingMRGPRX2, pseudo-allergic reactions, itch, pain, inflammatory orautoimmune disorders can be eased.

In some embodiments, the MRGPRX2 dependent condition is a condition thatis caused by IgE independent activation of MRGPRX2. IgE independentactivation of MRGPRX2 is capable of inducing mast cell degranulation.For example, IgE independent mast cell activation associated with somecases of chronic urticaria and other mast cell mediated conditions,which are not responsive to current anti-IgE or antihistamine therapies.Thus, the compounds of the present disclosure may be used for treatingan MRGPRX2 dependent condition caused by IgE independent activation ofMRGPRX2 and that would benefit from modulating MRGPRX2.

In some embodiments, the MRGPRX2 dependent condition is an itchassociated condition, a pain associated condition, a pseudo-allergicreaction, or an autoimmune or inflammatory disorder.

As used herein the phrase “pseudo-allergic reaction” refers to anIgE-independent allergic reaction, characterized by histamine release,inflammation, airway contraction, or any combination thereof. Apseudo-allergic reaction may be an anaphylactic reaction. Apseudo-allergic reaction may be caused by a range of cationicsubstances, collectively called basic secretagogues, includinginflammatory peptides and drugs associated with allergic-type reactions.Thus, in one embodiment, the method of present invention is provided totreat a pseudo-allergic reaction, such as pseudo-allergic reactionscaused by secretagogues, cationic peptidergic drugs, anionic peptidergicdrugs, neutral peptidergic drugs, non-steroidal antagonist drugs,neuropeptides, and antimicrobial peptides. In one embodiment, thepseudo-allergic reaction is caused by MCD peptide, Substance P, VIP,PACAP, dynorphin, somatostatin, Compound 48/80, cortistatin-14,mastoparan, melettin, cathelicidin peptides, ciprofloxacin, vancomycin,leuprolide, goserelin, histrelin, triptorelin, cetrorelix, ganirelix,degarelix, octreotide, lanreotide, pasireotide, sermorelin, tesamorelin,icatibant, glatiramer acetate, teriparatide, pramlintide, bleomycin,exenatide, glucagon, liraglutide, enfuvirtide, colistimethate,succinylcholine, tubocurarine, atracurium, mivacurium, and rocuronium.

As used herein, the phrase “itch associated condition” means pruritus(including acute and chronic pruritus) associated with any condition.The itch sensation can originate, e.g., from the peripheral nervoussystem (e.g., dermal or neuropathic itch) or from the central nervoussystem (e.g., neuropathic, neurogenic or psychogenic itch). Thus, in oneembodiment, the method of present invention is provided to treat an itchassociated condition, such as chronic itch; contact dermatitis; Allergicblepharitis; Anaphylaxis; Anaphylactoid drug reactions; Anaphylacticshock; Anemia; Atopic dermatitis; Bullous pemphigoid; Candidiasis;Chicken pox; end-stage renal failure; hemodialysis; Cholestaticpruritis; Chronic urticaria; Contact dermatitis, Dermatitisherpetiformis; Diabetes; Drug allergy, Dry skin; Dyshidrotic dermatitis;Ectopic eczema; Eosinophilic fasciitis; Epidermolysis bullosa;Erythrasma; Food allergy; Folliculitis; Fungal skin infection;Hemorrhoids; Herpes; HIV infection; Hodgkin's disease; Hyperthyroidism;Iodinated contrast dye allergy; Iron deficiency anemia; Kidney disease;Leukemia, porphyrias; Lymphoma; Mast cell activation syndrome,Malignancy; Mastocystosis; Multiple myeloma; Neurodermatitis;Onchocerciasis; Paget's disease; Pediculosis; Polycythemia rubra vera;Prurigo nodularis; Lichen Planus; Lichen Sclerosis; Pruritus ani;Pseudo-allergic reactions; Pseudorabies; Psoriasis; Rectal prolapse;Sarcoidosis granulomas; Scabies; Schistosomiasis; Scleroderma, Severestress, Stasia dermatitis; Swimmer's itch; Thyroid disease; Tineacruris; Uremic Pruritus; Rosacea; Cutaneous amyloidosis; Scleroderma;Acne; wound healing; burn healing; ocular itch; and Urticaria.

As used herein, the phrase “pain associated condition” means any paindue to a medical condition. Thus, in one embodiment, the method ofpresent invention is provided to treat a pain associated condition, suchas Acute Pain, Advanced Prostate Cancer, AIDS-Related Pain, AnkylosingSpondylitis, Arachnoiditis, Arthritis, Arthrofibrosis, Ataxic CerebralPalsy, Autoimmune Atrophic Gastritis, Avascular Necrosis, Back Pain,Behcet's Disease (Syndrome), Burning Mouth Syndrome, Bursitis, CancerPain, Carpal Tunnel, Cauda Equina Syndrome, Central Pain Syndrome,Cerebral Palsy, Cervical Stenosis, Charcot-Marie-Tooth (CMT) Disease,Chronic Fatigue Syndrome (CFS), Chronic Functional Abdominal Pain(CFAP), Chronic Pain, Chronic Pancreatitis, Chronic Pelvic PainSyndrome, Collapsed Lung (Pneumothorax), Complex Regional Pain Syndrome(RSD), Corneal Neuropathic Pain, Crohn's Disease, Degenerative DiscDisease, Dental Pain, Dercum's Disease, Dermatomyositis, DiabeticPeripheral Neuropathy (DPN), Dystonia, Ehlers-Danlos Syndrome (EDS),Endometriosis, Eosinophilia-Myalgia Syndrome (EMS), Erythromelalgia,Fibromyalgia, Gout, Headaches, Herniated disc, Hydrocephalus,Intercostal Neuraligia, Interstitial Cystitis, Irritable Bowel syndrome(IBS), Juvenile Dermatositis (Dermatomyositis), Knee Injury, Leg Pain,Loin Pain-Haematuria Syndrome, Lupus, Lyme Disease, Medullary SpongeKidney (MSK), Meralgia Paresthetica, Mesothelioma, Migraine,Musculoskeletal pain, Myofascial Pain, Myositis, Neck Pain, NeuropathicPain, Occipital Neuralgia, Osteoarthritis, Paget's Disease, ParsonageTurner Syndrome, Pelvic Pain, Periodontitis Pain, Peripheral Neuropathy,Phantom Limb Pain, Pinched Nerve, Polycystic Kidney Disease, PolymyalgiaRhuematica, Polymyositis, Porphyria, Post Herniorraphy Pain Syndrome,Post Mastectomy, Postoperative Pain, Pain Syndrome, Post Stroke Pain,Post Thorocotomy Pain Syndrome, Postherpetic Neuralgia (Shingles),Post-Polio Syndrome, Primary Lateral Sclerosis, Psoriatic Arthritis,Pudendal Neuralgia, Radiculopathy, Raynaud's Disease, RheumatoidArthritis (RA), Sacroiliac Joint Dysfunction, Sarcoidosi, Scheuemann'sKyphosis Disease, Sciatica, Scoliosis, Shingles (Herpes Zoster),Sjogren's Syndrome, Spasmodic Torticollis, Sphincter of OddiDysfunction, Spinal Cerebellum Ataxia (SCA Ataxia), Spinal Cord Injury,Spinal Stenosis, Syringomyelia, Tarlov Cysts, Transverse Myelitis,Trigeminal Neuralgia, Neuropathic Pain, Ulcerative Colitis, VascularPain and Vulvodynia.

As used herein, the term “autoimmune disorder”, or “inflammatorydisorder” means a disease or disorder arising from and/or directedagainst an individual's own tissues or organs, or a co-segregate ormanifestation thereof, or resulting condition therefrom. Typically,various clinical and laboratory markers of autoimmune diseases may existincluding, but not limited to, hypergammaglobulinemia, high levels ofautoantibodies, antigen-antibody complex deposits in tissues, clinicalbenefit from corticosteroid or immunosuppressive treatments, andlymphoid cell aggregates in affected tissues. Thus, in one embodiment,the method of present invention is provided to treat an autoimmunedisorder, such as chronic inflammation, mast cell activation syndrome,Multiple Sclerosis, Steven Johnson's Syndrome, Toxic EpidermalNecrolysis, appendicitis, bursitis, cutaneous lupus, colitis, cystitis,dermatitis, phlebitis, reflex sympathetic dystrophy/complex regionalpain syndrome (rsd/crps), rhinitis, tendonitis, tonsillitis, acnevulgaris, sinusitis, rosacea, psoriasis, graft-versus-host disease,reactive airway disorder, asthma, airway infection, allergic rhinitis,autoinflammatory disease, celiac disease, chronic prostatitis,diverticulitis, glomerulonephritis, hidradenitis suppurativa,hypersensitivities, intestinal disorder, epithelial intestinal disorder,inflammatory bowel disease, irritable bowel syndrome, Crohn's Disease,ulcerative colitis, lupus erythematous, interstitial cystitis, otitis,pelvic inflammatory disease, endometrial pain, reperfusion injury,rheumatic fever, rheumatoid arthritis, sarcoidosis, transplantrejection, psoriasis, lung inflammation, chronic obstructive pulmonarydisease, permanent sputum eosiniophilia, eosinophilic leukemia,eosinophilic esophagitis, eosinophilic gastritis, mast cellgastrointestinal disease, hypereosinophilic syndrome,aspirin-exacerbated respiratory disease, nasal polyposis, chronicrhinosinusitis, antibody-dependent cell-mediated cytotoxicity,neurofibromatosis, swannamatoisis, tubulointerstitial nephritis,glomerulonephritis, diabetic nephropathy, allograft rejection,amyloidosis, renovascular ischemia, reflux nephropathy, polycystickidney disease, liver fibrosis/cirrhosis, autoimmune liver disease,Biliary atresia, acute and chronic Hepatitis B and C virus, Liver tumorsand cancer, Alcoholic liver disease, Polycystic liver disease, Livercholangiocarcinoma, neuromyelitis optica spectum disorder,cardiovascular disease, and vasculitis.

As used herein the phrase “cancer associated condition” means anydisease arising from the proliferation of malignant cancerous cells.Thus, in one embodiment, the method of present invention is provided totreat a cancer/tumor associated condition, such as adenoid cysticcarcinoma, adrenal gland tumor, amyloidosis, anal cancer, appendixcancer, astrocytoma, ataxia-telangiectasia, beckwith-wiedemann syndrome,cholangiocarcinoma, birt-hogg-dube syndrome, bone cancer, brain stemglioma, brain tumor, breast cancer (inflammatory, metastatic, male),prostrate, basal cell, melanoma, colon, colorectal, bladder, kidneycancer, lacrimal gland cancer, laryngeal and hypopharyngeal cancer, lungcancer (non-small cell, small cell), leukemia (acute lymphoblastic,acute lymphocytic, acute myeloid, B cell prolymphocytic, chroniclymphocytic, chronic myeloid, chronic T cell lymphocytic, eosinophilic),Liver Cancer, Li-Fraumei syndrome, lymphoma (Hodgkin and non-hodgkin),lynch syndrome, mastocytosis, medulloblastoma, meningioma, mesothelioma,multiple endocrine neoplasia, multiple myeloma, MUTYH-associatedpolyposis, myelodyspastic syndrome, nasal cavity and paranasal sinuscancer, neurobastoma, neuroendocrine tyymors, neurofibromatosis, penilecancer, parathyroid cancer, ovarian fallopian tube and peritonealcancer, osteosarcoma, pituitary gland tumor, pleupulmonary blastoma,oral and oropharyngeal, thyroid, uterine, pancreatic, carney complex,brain and spinal cord cancer, cervical cancer, cowden syndrome,craniopharyngioma, desmoid tumor, desmoplatic infantile ganglioglioma,ependymoma, esophageal cancer, ewing sarcoma, eye cancer, eyelid cancer,familial adenomatous polyposis, familial GIST, familial malignantmelanoma, familial pancreatic cancer, gallbladder cancer,gastrointestinal stromal tumor, germ cell tumor, gestationaltrophoblastic disease, head and neck cancer, hereditary breast andovarian cancer, hereditary diffuse gastric cancer, hereditary,leiomyomastosis and renal cell cancer, hereditary pancreatitis,herediatary papillary renal carcinoma, hereditary mixed polyposissyndrome, HIV/AIDS related cancers, retinoblastoma, rhabdomyosarcoma,salivary glanc cancer, Kaposi sarcoma, small bowel cancer, stomachcancer, testicular cancer, thymoma and thymic carcinoma, thyroid cancer,vaginal cancer, culver cancer, werner syndrome and Xerodermapigmentosum.

As used herein, the term “administration” refers to providing acompound, or a pharmaceutical composition comprising the compound asdescribed herein. The compound or composition can be administered byanother person to the subject or it can be self-administered by thesubject. Non-limiting examples of routes of administration are oral,parenteral (e.g., intravenous), or topical.

As used herein, the term “treatment” refers to an intervention thatameliorates a sign or symptom of a disease or pathological condition. Asused herein, the terms “treatment”, “treat” and “treating,” withreference to a disease, pathological condition or symptom, also refersto any observable beneficial effect of the treatment. The beneficialeffect can be evidenced, for example, by a delayed onset of clinicalsymptoms of the disease in a susceptible subject, a reduction inseverity of some or all clinical symptoms of the disease, a slowerprogression of the disease, a reduction in the number of relapses of thedisease, an improvement in the overall health or well-being of thesubject, or by other parameters well known in the art that are specificto the particular disease. A prophylactic treatment is a treatmentadministered to a subject who does not exhibit signs of a disease orexhibits only early signs, for the purpose of decreasing the risk ofdeveloping pathology. A therapeutic treatment is a treatmentadministered to a subject after signs and symptoms of the disease havedeveloped.

As used herein, the term “subject” refers to an animal (e.g., a mammal,such as a human, dog or horse). A subject to be treated according to themethods described herein may be one who has been diagnosed with aMRGPRX2 dependent condition or MRGPRX2 ortholog B2 dependent condition,such as a pseudo-allergic reaction, an itch associated condition, a painassociated condition, inflammatory or an autoimmune disorder. Diagnosismay be performed by any method or technique known in the art. Oneskilled in the art will understand that a subject to be treatedaccording to the present disclosure may have been subjected to standardtests or may have been identified, without examination, as one at riskdue to the presence of one or more risk factors associated with thedisease or condition. The term “patient” may be used interchangeablywith the term “subject.” A subject may refer to an adult or pediatricsubject.

The Federal Food, Drug, and Cosmetic Act defines “pediatric” as asubject aged 21 or younger at the time of their diagnosis or treatment.Pediatric subpopulations are further characterized as: (i) neonates—frombirth through the first 28 days of life; (ii) infants—from 29 days toless than 2 years; (iii) children—2 years to less than 12 years; and(iv) adolescents—aged 12 through 21. Despite the definition, dependingon the susceptible patient population and clinical trial evaluation, anapproved regulatory label may include phrasing that specificallymodifies the range of a pediatric population, such as, for example,pediatric patients up to 22 years of age.

In another embodiment, the method of treating a subject having a MRGPRX2dependent condition (e.g., an itch associated condition, a painassociated condition, a pseudo-allergic reaction, or an inflammatory orautoimmune disorder) described herein further comprises administering tothe subject a pharmaceutically effective amount of a second therapeuticagent. In one embodiment, the itch associated condition is apseudo-allergic condition.

In one embodiment, the second therapeutic agent is an antihistamine,such as an H1 receptor antagonist or an H2 receptor antagonist. In oneembodiment, the second therapeutic agent is an H1 receptor antagonistantihistamine, such as levocetirizine, loratadine, fexofenadine,cetirizine, desloratadine, olopatadine, diphenhydramine, cyproheptadineor hydroxyzine pamoate. In one embodiment, the second therapeutic agentis a H2 receptor antagonist, such as cimetidine, nizatidine, ranitidineor famotidine. In one embodiment, the second therapeutic agent is aleukotriene receptor antagonist or leukotriene synthesis inhibitor, suchas montelukast, zafirlukast, pranlukast, or 5-lipoxygenase inhibitor(e.g., zileuton, hypericum perforatum). In one embodiment, the secondtherapeutic agent is an immunomodulatory agent such as Omalizumab orimmunoglobulin therapy. In one embodiment, the second therapeutic agentis a corticosteroid, such as hydrocortisone, cortisone, ethamethasoneb,triamcinolone, prednisone, prednisolone, or fludrocortisone. In oneembodiment, the second therapeutic agent is a tricylic antidepressantthat can relieve itch such as doxepin, amitriptyline or nortriptyline.In one embodiment, the second therapeutic agent is an anti-inflammatorydrug such as dapsone, sulfasalazine, hydroxycholoroquine or colchicine.In one embodiment, the second therapeutic agent is an immunosuppressantsuch as cyclosporine, methotrexate, mycophenolic acid or tacromilus.

The second therapeutic agent may be administered simultaneously,separately, or sequentially with the compounds of the presentdisclosure. If administered simultaneously, the second therapeutic agentand compound of the present disclosure may be administered in separatedosage forms or in the same dosage form.

In another embodiment, a method of treating a subject having an itchassociated condition is provided, the method comprising administering tothe subject a pharmaceutically effective amount of a compound havingstructure (I) or a pharmaceutically acceptable salt, isomer, hydrate,solvate or isotope thereof, or a pharmaceutical composition thereof. Inone embodiment, the itch associated condition is urticaria, pruritus,atopic dermatitis, dry skin, psoriasis, contact dermatitis, or eczema.In another embodiment, a method of treating a subject having aninflammation or autoimmune associated condition is provided, the methodcomprising administering to the subject a pharmaceutically effectiveamount of a compound having structure (I) or a pharmaceuticallyacceptable salt, isomer, hydrate, solvate or isotope thereof, or apharmaceutical composition thereof. In one embodiment, the inflammationor autoimmune associated condition is sinusitis, asthma, rosacea, orendometriosis.

In another embodiment, a method of treating a subject having a painassociated condition is provided, the method comprising administering tothe subject a pharmaceutically effective amount of a compound havingstructure (I) or a pharmaceutically acceptable salt, isomer, hydrate,solvate or isotope thereof, or a pharmaceutical composition thereof. Inone embodiment, the pain associated condition is chronic pelvic painsyndrome, endometriosis pain, fibromyalgia, migraine or postoperativepain.

Compounds

As detailed above, the present disclosure provides compounds showingsignificant activity as MRGPRX2 antagonists. Accordingly, one embodimentprovides a compound having the following structure (I):

or a pharmaceutically acceptable salt, isomer, hydrate, solvate orisotope thereof, wherein:

R¹ is cycloalkyl, aryl, heterocyclyl, —(CH₂)_(n)Q, —CHQR, or —CQ(R)₂,wherein R¹ is optionally substituted with one or more R^(q1);

Q is C₁₋₆ alkyl, aryl, cycloalkyl, heterocyclyl, —CH₂C(O)OR, —C(O)OR,—C(O)NHR, trihalomethyl, dihalomethyl, —CN, —N(R)₂, —N(R)C(O)R,—N(R)C(O)OR, or —N(R)S(O)₂R, wherein Q is optionally substituted withone or more R^(q2);

R^(q1) and R^(q2) are independently H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl, aryl, cycloalkyl, heteroaryl, heterocyclyl, —OR, —O(CH₂)_(n)R,haloalkoxy, —C(O)OR, —C(O)R, —OC(O)R, halo, haloalkyl, —CN, —N(R)₂,—N(R)C(═NH)N(R)₂, —N(R)C(O)R, —N(R)S(O)₂R, S(O)₂R, —C(H)Q′R, or—(CH₂)_(n)Q′ where Q′ is selected from C₁₋₆ alkyl, aryl, cycloalkyl,heterocyclyl, OR′, —C(O)OR′, —OC(O)R′, haloalkyl, —CN, —N(R′)₂,—N(R′)C(O)R′, and —N(R′)S(O)₂R′;

D is N or CR^(d);

W is N or CR^(w);

Z is N or CR^(z);

each R is independently H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,—C(O)NHR′, amino, —(CH₂)_(n)R′, halo, aryl, cycloalkyl, heteroaryl orheterocyclyl, or two R groups taken together with the atom to which theyare attached form a carbocyle or heterocycle;

R′ is H, C₁₋₆ alkyl, haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl,cycloalkyl, heteroaryl, or heterocyclyl;

R^(d) is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, —CN, halo, —C(O)R,aryl, cycloalkyl, heteroaryl, or heterocyclyl;

R^(w) is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, —CN, halo, —C(O)R,aryl, cycloalkyl, heteroaryl, or heterocyclyl;

R^(z) is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, —CN, halo, —C(O)R,aryl, cycloalkyl, heteroaryl, or heterocyclyl;

each n is independently 0, 1, 2, 3, 4 or 5;

when D is CR^(d)′, W is CR^(w), and Z is CR^(z), then R² is H, C₁₋₆alkyl, C₂₋₆ alkenyl, cycloalkyl, —OR, —C(O)OR, —OC(O)R, halo, —CF₃,—CF₂H, —C(F)H₂, C(F)₂R, —C(F)(R)₂, —N(R)₂, —N(R)C(O)R, —N(R)S(O)₂R, orS(O)₂R; or

when at least one of D, W or Z is N, then R² is C₁₋₆ alkyl, C₂₋₆alkenyl, cycloalkyl, —OR, —C(O)OR, —OC(O)R, halo, —CF₃, —CF₂H, —C(F)H₂,C(F)₂R, —C(F)(R)₂, —N(R)₂, —N(R)C(O)R, —N(R)S(O)₂R, or S(O)₂R;

each R³ is independently H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,aryl, cycloalkyl, heteroaryl, heterocyclyl, —OR, —C(O)OR, —OC(O)R, halo,haloalkyl, —CN, —N(R)₂, —N(R)C(O)R, —N(R)S(O)₂R, or S(O)₂R;

with the provisos that:

-   -   when D is CR^(d), then W and Z cannot both be N;    -   when D and Z are both N, then R² cannot be dimethylamino; and    -   when D is CH, Z is CH and W is CR^(w), then R^(w) is not cyano        or piperidine.

In another embodiment, are provided compounds of the followingstructures (Ia) or (Ia′):

or a pharmaceutically acceptable salt, isomer, hydrate, solvate orisotope thereof.

In yet another embodiment, are provided compounds of the followingstructures (Ib), (Ic), (Id), (Ie), (If) or (Ig):

or a pharmaceutically acceptable salt, isomer, hydrate, solvate orisotope thereof.

In one embodiment R¹ is aryl, —(CH₂)_(n)Q, —CHQR, or —CQ(R)₂, wherein R¹is optionally substituted with one or more R^(q1);

Q is aryl, —CH₂C(O)OR, —C(O)OR, —C(O)NHR, haloalkyl, —CN, —N(R)₂,—N(R)C(O)R, —N(R)C(O)OR, or —N(R)S(O)₂R, wherein Q is optionallysubstituted with one or more R^(q2).

In a specific embodiment, R¹ is phenyl.

In some embodiments, the aryl is unsubstituted.

In yet other embodiments, the aryl is substituted with one or more R. Insome other embodiments, R^(q1) is H, —OR, —N(R)₂, —N(R)S(O)₂R, —CN,—N(R)C(O)R, —C(H)Q′R, heterocyclyl, or halo.

In one embodiment, R¹ is heterocyclyl, —(CH₂)_(n)Q, —CHQR, or —CQ(R)₂,wherein R¹ is optionally substituted with one or more R^(q1);

Q is heterocyclyl, —CH₂C(O)OR, —C(O)OR, —C(O)NHR, haloalkyl, —CN,—N(R)₂, —N(R)C(O)R, —N(R)C(O)OR, or —N(R)S(O)₂R, wherein Q is optionallysubstituted with one or more R^(q2).

In some embodiments, the heterocyclyl is unsubstituted.

In one embodiment, the heterocyclyl is substituted with one or moreR^(q1). In some other embodiments, R^(q1) is H, C₁₋₆ alkyl, —N(R)₂, orcycloalkyl.

In one embodiment, R¹ is —(CH₂)_(n)Q;

Q is —N(R)₂, —N(R)C(O)R, —N(R)C(O)OR, or —N(R)S(O)₂R, wherein Q isoptionally substituted with one or more R^(q2).

In a specific embodiment, n is 0 and Q is N(R)₂.

In yet other embodiments, R¹ is —CQ(R)₂;

Q is—trihalomethyl, —CN, —N(R)₂, —N(R)C(O)R, —N(R)C(O)OR, or—N(R)S(O)₂R, wherein Q is optionally substituted with one or more V.

In a specific embodiment, Q is trihalomethyl.

In one embodiment, each R³ is H.

In another embodiment, D is CR^(d), W is CR^(w) and Z is CR^(z) andwherein each R^(d), R^(w) and R^(z) are independently C₁₋₆ alkyl, H,—CN, halo, or —C(O)R.

In some embodiments, D is N, W is CR^(w) and Z is CR^(z) and wherein R²is —CF₃, C₁₋₆ alkyl, —N(R)₂, cycloalkyl, —C(O)OR, or halo.

In a specific embodiment, R² is —CF₃.

In another embodiment, R² is C₁₋₆ alkyl.

In yet another embodiment, R² is —N(R)₂.

In some other embodiments, R² is cycloalkyl.

In yet another embodiment, R² is —C(O)OR.

In one embodiment, R² is halo.

In some other embodiments, at least two of D, W and Z is N, and whereinR² is C₁₋₆ alkyl, —OR, or —N(R)₂.

In one embodiment, a compound is selected from any one of the compoundslisted in Table I, or a pharmaceutically acceptable salt, isomer,hydrate, solvate or isotope thereof.

TABLE I Representative compounds having Structure (I) Structure Cpd No.

1-1 

1-2 

1-3 

1-4 

1-5 

1-6 

1-7 

1-8 

1-9 

1-10

1-11

1-12

1-13

1-14

1-15

1-16

1-17

1-18

1-19

1-20

1-21

1-22

1-23

1-24

1-25

2-1 

2-2 

2-3 

3-1 

4-1 

4-2 

4-3 

4-4 

4-5 

4-6 

4-7 

4-8 

4-9 

4-10

4-11

4-12

4-13

4-14

4-15

4-16

4-17

4-18

4-19

4-20

4-21

4-22

4-23

4-24

4-25

4-26

4-27

4-28

4-29

4-30

4-31

4-32

4-33

4-34

4-35

4-36

4-37

4-38

4-39

4-40

4-41

4-42

4-43

4-44

4-45

4-46

4-47

4-48

4-49

4-50

4-51

4-52

4-53

4-54

4-55

4-56

4-57

4-58

4-59

4-60

4-61

4-62

4-63

4-64

4-65

4-66

5-1 

5-2 

5-3 

5-4 

5-5 

5-6 

5-7 

5-8 

5-9 

5-10

Representative compounds of structure (I), as well as structures (Ia),(Ia′), (Ib), (Ic), (Id), (Ie), (If) or (Ig) as applicable, include, butnot limited to, any one of the compounds listed below in their IUPACnames as well as a pharmaceutically acceptable salt, isomer, hydrate,solvate or isotope thereof.

-   N-[(1R,3S)-3-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-4-methoxybenzamide;-   N-[(1R,3S)-3-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-methoxy-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   4-methoxy-N-[(1R,3S)-3-{[2-(trifluoromethyl)-5H,6H,7H-cyclopenta[d]pyrimidin-4-yl]amino}cyclohexyl]benzamide;-   N-[(1s,4s)-4-{[2-(dimethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino}cyclohexyl]benzamide;-   4-fluoro-N-[(1s,4s)-4-{[2-(pyrrolidin-1-yl)-6-(trifluoromethyl)pyrimidin-4-yl]amino}cyclohexyl]benzamide;-   4-fluoro-N-[(1s,4s)-4-{[2-(dimethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino}cyclohexyl]benzamide;-   4-fluoro-N-[(1s,4s)-4-{[6-methoxy-2-(trifluoromethyl)pyrimidin-4-yl]amino}cyclohexyl]benzamide;-   4-(dimethylamino)-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   3-methyl-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]furan-2-carboxamide;-   3-cyano-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-fluoro-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-fluoro-N-[(1s,4s)-4-{[2-(trifluoromethyl)-5,6,7,8-tetrahydroquinazolin-4-yl]amino}cyclohexyl]benzamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-indole-4-carboxamide;-   4-(dimethylamino)-2-fluoro-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   4-(propane-2-sulfonamido)-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   2-amino-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1,3-thiazole-4-carboxamide;-   3-(dimethylamino)-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   1-methyl-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrazole-5-carboxamide;-   4-(N-methylmethanesulfonamido)-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-indole-3-carboxamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]imidazo[1,2-a]pyridine-6-carboxamide;-   4-(dimethylamino)-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   4-fluoro-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   3-methoxy-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1,3-benzothiazole-7-carboxamide;-   4-(morpholin-4-yl)-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   N-[(1R,3S)-3-{[3,5-bis(trifluoromethyl)phenyl]amino}cyclohexyl]-4-methoxybenzamide;-   4-fluoro-N-[(1s,4s)-4-{[3,5-bis(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   4-fluoro-N-[(1s,4s)-4-{[2-chloro-6-(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-fluoro-N-[(1s,4s)-4-{[2-cyclopropyl-6-(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-methoxy-N-[(1s,4s)-4-({2-[(3-methylphenyl)amino]-6-(trifluoromethyl)pyridin-4-yl}amino)cyclohexyl]benzamide;-   4-fluoro-N-[(1s,4s)-4-{[2-(phenylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino}cyclohexyl]benzamide;-   4-(dimethylamino)-N-[(1s,4s)-4-({2-[ethyl(methyl)amino]-6-(trifluoromethyl)pyrimidin-4-yl}amino)cyclohexyl]benzamide;-   4-methoxy-N-[(1s,4s)-4-{[2-(ethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino}cyclohexyl]benzamide;-   4-(dimethylamino)-N-[(1s,4s)-4-{[2-(dimethylamino)-6-(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-methoxy-N-[(1s,4s)-4-{[4-fluoro-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   3-(dimethylamino)-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-methoxy-N-[(1s,4s)-4-{[2-(dimethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino}cyclohexyl]benzamide;-   4-methoxy-N-[(1s,4s)-4-({2-[methyl(propan-2-yl)amino]-6-(trifluoromethyl)pyrimidin-4-yl}amino)cyclohexyl]benzamide;-   4-methoxy-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-(dimethylamino)-N-[(1s,4s)-4-{[2-(dimethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino}cyclohexyl]benzamide;-   3-cyano-N-[(1s,4s)-4-{[2-(dimethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino}cyclohexyl]benzamide;-   6-methyl-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1H-1,3-benzodiazole-5-carboxamide;-   2,3-dioxo-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-2,3-dihydro-1H-indole-7-carboxamide;-   1-ethyl-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1H-pyrazole-5-carboxamide;-   4-cyano-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-chloro-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   1-methyl-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1,2,3,4-tetrahydroquinoline-7-carboxamide;-   3-(2-methylpropanamido)-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   3-(1-cyanoethyl)-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   3-methanesulfonamido-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1,3-benzothiazole-7-carboxamide;-   N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1H-indole-4-carboxamide;-   3-acetamido-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-(methylamino)-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   3-[(methylcarbamoyl)amino]-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1H-pyrrole-1-carboxamide;-   3-methoxy-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   1-methyl-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1,2,3,4-tetrahydroquinoline-5-carboxamide;-   4-carbamimidamido-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   1-methyl-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1,2,3,4-tetrahydroquinoline-6-carboxamide;-   N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1H-indole-3-carboxamide;-   4-(N-methylmethanesulfonamido)-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-(3,5-dimethyl-1H-pyrazol-1-yl)-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-(1H-imidazol-1-yl)-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   1-methyl-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1,2,3,4-tetrahydroquinoline-8-carboxamide;-   2-amino-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1,3-thiazole-4-carboxamide;-   3-hydroxy-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   3,3-dimethyl-1-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]urea;-   N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-4-(1H-1,2,3,4-tetrazol-1-yl)benzamide;-   4-(1H-pyrazol-1-yl)-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   2,4-dichloro-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   4-(3-methyl-5-oxo-4,5-dihydro-1H-pyrazol-1-yl)-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   2-methyl-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1,2,3,4-tetrahydroisoquinoline-5-carboxamide;-   N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-1H-1,3-benzodiazole-2-carboxamide;-   4-methyl-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]piperazine-1-carboxamide;-   N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-4,5,6,7-tetrahydro-1H-indole-2-carboxamide;-   4-(1,1-dioxo-1λ⁶-thiomorpholin-4-yl)-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]imidazo[1,2-a]pyridine-6-carboxamide;-   1-methyl-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]pyrrolidine-3-carboxamide;-   4-(pyrrolidin-1-yl)-N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   N-[(1s,4s)-4-{[2,6-bis(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]-5,6,7,8-tetrahydronaphthalene-2-carboxamide;-   4-methoxy-N-[(1s,4s)-4-{[2-(difluoromethyl)-6-(trifluoromethyl)pyridin-4-yl]amino}cyclohexyl]benzamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxamide;-   N-[(1s,4s)-4-{[4-cyano-3-methyl-5-(trifluoromethyl)phenyl]amino}cyclohexyl]imidazo[1,2-a]pyridine-6-carboxamide;-   5-chloro-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrazole-4-carboxamide;-   5-cyclopropyl-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrazole-4-carboxamide;-   4-(methylamino)-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   4-acetamido-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-indazole-7-carboxamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-indazole-4-carboxamide;-   4-(4-methyl-1H-imidazol-1-yl)-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrrolo[2,3-b]pyridine-4-carboxamide;-   4-[(2,2-difluoroethyl)amino]-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   4-[(2,2-difluoroethyl)(methyl)amino]-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   2-[(propan-2-yl)amino]-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]pyridine-4-carboxamide;-   5-cyano-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrazole-4-carboxamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrrolo[2,3-b]pyridine-3-carboxamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrazolo[4,3-b]pyridine-7-carboxamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrazolo[3,4-b]pyridine-4-carboxamide;-   2-methyl-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrrolo[2,3-b]pyridine-4-carboxamide;-   1-methyl-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrrolo[2,3-b]pyridine-4-carboxamide;-   4-methyl-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-indole-3-carboxamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrrolo[3,2-c]pyridine-3-carboxamide;-   N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrrolo[2,3-c]pyridine-3-carboxamide;-   N1-methyl-N4-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]benzene-1,4-dicarboxamide;-   2-(methylamino)-N-[(1s,4s)-4-{[4-cyano-3-(trifluoromethyl)phenyl]amino}cyclohexyl]pyridine-4-carboxamide;-   2,6-dimethyl-N-[(1s,4s)-4-{[4-cyano-3-methyl-5-(trifluoromethyl)phenyl]amino}cyclohexyl]pyridine-4-carboxamide;-   2-[(propan-2-yl)amino]-N-[(1s,4s)-4-{[4-cyano-3-methyl-5-(trifluoromethyl)phenyl]amino}cyclohexyl]pyridine-4-carboxamide;-   4-(methylamino)-N-[(1s,4s)-4-{[4-cyano-3-methyl-5-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   2-(methylamino)-N-[(1s,4s)-4-{[4-cyano-3-methyl-5-(trifluoromethyl)phenyl]amino}cyclohexyl]pyridine-4-carboxamide;-   N-[(1s,4s)-4-{[4-cyano-3-methyl-5-(trifluoromethyl)phenyl]amino}cyclohexyl]-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxamide;-   1-ethyl-N-[(1s,4s)-4-{[4-cyano-3-methyl-5-(trifluoromethyl)phenyl]amino}cyclohexyl]-1H-pyrazole-5-carboxamide;-   4-[(2,2-difluoroethyl)(methyl)amino]-N-[(1s,4s)-4-{[4-cyano-3-methyl-5-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;-   N-((1s,4s)-4-((4-cyano-3-(trifluoromethyl)phenyl)amino)cyclohexyl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxamide;-   4-[(2,2-difluoroethyl)amino]-N-[(1s,4s)-4-{[4-cyano-3-methyl-5-(trifluoromethyl)phenyl]amino}cyclohexyl]benzamide;    and-   N-[(1s,4s)-4-{[4-cyano-3-methyl-5-(trifluoromethyl)phenyl]amino}cyclohexyl]imidazo[1,5-a]pyridine-6-carboxamide.

Pharmaceutical Compositions

In certain embodiments, the invention provides a pharmaceuticalcomposition comprising a compound of structure (I) or any of thestructures (Ia), (Ia′), (Ib), (Ic), (Id), (Ie), (If) or (Ig), or apharmaceutically acceptable salt, isomer, hydrate, solvate or isotopethereof, together with at least one pharmaceutically acceptable carrier,diluent, or excipient. For example, the active compound will usually bemixed with a carrier, or diluted by a carrier, or enclosed within acarrier which can be in the form of an ampoule, capsule, sachet, paper,or other container. When the active compound is mixed with a carrier, orwhen the carrier serves as a diluent, it can be solid, semi-solid, orliquid material that acts as a vehicle, excipient, or medium for theactive compound. The active compound can be adsorbed on a granular solidcarrier, for example contained in a sachet. Some examples of suitablecarriers are water, salt solutions, alcohols, polyethylene glycols,polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatin,lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugar,cyclodextrin, amylose, magnesium stearate, talc, gelatin, agar, pectin,acacia, stearic acid, or lower alkyl ethers of cellulose, silicic acid,fatty acids, fatty acid amines, fatty acid monoglycerides anddiglycerides, pentaerythritol fatty acid esters, polyoxyethylene,hydroxymethylcellulose, and polyvinylpyrrolidone. Similarly, the carrieror diluent can include any sustained release material known in the art,such as glyceryl monostearate or glyceryl distearate, alone or mixedwith a wax.

As used herein, the term “pharmaceutical composition” refers to acomposition containing one or more of the compounds described herein, ora pharmaceutically acceptable isomer, racemate, hydrate, solvate,isotope or salt thereof, formulated with a pharmaceutically acceptablecarrier, which can also include other additives, and manufactured orsold with the approval of a governmental regulatory agency as part of atherapeutic regimen for the treatment of disease in a mammal.Pharmaceutical compositions can be formulated, for example, for oraladministration in unit dosage form (e.g., a tablet, capsule, caplet,gelcap, or syrup); for topical administration (e.g., as a cream, gel,lotion, or ointment); for intravenous administration (e.g., as a sterilesolution free of particulate emboli and in a solvent system suitable forintravenous use); for administration to a pediatric subject (e.g.,solution, syrup, suspension, elixir, powder for reconstitution assuspension or solution, dispersible/effervescent tablet, chewabletablet, lollipop, freezer pops, troches, oral thin strips, orallydisintegrating tablet, orally disintegrating strip, and sprinkle oralpowder or granules); or in any other formulation described herein.Conventional procedures and ingredients for the selection andpreparation of suitable formulations are described, for example, inRemington: The Science and Practice of Pharmacy, 21st Ed., Gennaro, Ed.,Lippencott Williams & Wilkins (2005) and in The United StatesPharmacopeia: The National Formulary (USP 36 NF31), published in 2013.

In some embodiments, the pharmaceutical composition comprising acompound of structure (I) or any of the structures (Ia), (Ia′), (Ib),(Ic), (Id), (Ie), (If) or (Ig), or a pharmaceutically acceptable salt,isomer, hydrate, solvate or isotope thereof, with at least onepharmaceutically acceptable carrier, diluent, or excipient furthercomprises a second therapeutic agent.

In one embodiment, the second therapeutic agent is an antihistamine,such as an H1 receptor antagonist or an H2 receptor antagonist. In oneembodiment, the second therapeutic agent is an H1 receptor antagonistantihistamine, such as levocetirizine, loratadine, fexofenadine,cetirizine, desloratadine, olopatadine, diphenhydramine, cyproheptadineor hydroxyzine pamoate. In one embodiment, the second therapeutic agentis a H2 receptor antagonist, such as cimetidine, nizatidine, ranitidineor famotidine. In one embodiment, the second therapeutic agent is aleukotriene receptor antagonist or leukotriene synthesis inhibitor, suchas montelukast, zafirlukast, pranlukast, or 5-lipoxygenase inhibitor(e.g., zileuton, hypericum perforatum). In one embodiment, the secondtherapeutic agent is an immunomodulatory agent such as Omalizumab orimmunoglobulin therapy. In one embodiment, the second therapeutic agentis a corticosteroid, such as hydrocortisone, cortisone, ethamethasoneb,triamcinolone, prednisone, prednisolone, or fludrocortisone. In oneembodiment, the second therapeutic agent is a tricylic antidepressantthat can relieve itch such as doxepin, amitriptyline or nortriptyline.In one embodiment, the second therapeutic agent is an anti-inflammatorydrug such as dapsone, sulfasalazine, hydroxycholoroquine or colchicine.In one embodiment, the second therapeutic agent is an immunosuppressantsuch as cyclosporine, methotrexate, mycophenolic acid or tacromilus.

In one embodiment, the second therapeutic agent is an H1 receptorantagonist antihistamine, such as levocetirizine, loratadine,fexofenadine, cetirizine, desloratadine, olopatadine, diphenhydramine,cyproheptadine or hydroxyzine pamoate. In one embodiment, the secondtherapeutic agent is a H2 receptor antagonist, such as cimetidine,nizatidine, ranitidine or famotidine. In one embodiment, the secondtherapeutic agent is a leukotriene receptor antagonist or leukotrienesynthesis inhibitor, such as montelukast, zafirlukast, pranlukast, or5-lipoxygenase inhibitor (e.g., zileuton, hypericum perforatum). In oneembodiment, the second therapeutic agent is an immunomodulatory agentsuch as Omalizumab or immunoglobulin therapy. In one embodiment, thesecond therapeutic agent is a corticosteroid, such as hydrocortisone,cortisone, ethamethasoneb, triamcinolone, prednisone, prednisolone, orfludrocortisone. In one embodiment, the second therapeutic agent is atricylic antidepressant that can relieve itch such as doxepin,amitriptyline or nortriptyline. In one embodiment, the secondtherapeutic agent is an anti-inflammatory drug such as dapsone,sulfasalazine, hydroxycholoroquine or colchicine. In one embodiment, thesecond therapeutic agent is an immunosuppressant such as cyclosporine,methotrexate, mycophenolic acid or tacromilus.

As used herein, the term “pharmaceutically acceptable carrier” refers toany ingredient other than the disclosed compounds, or a pharmaceuticallyacceptable isomer, racemate, hydrate, solvate, isotope or salt thereof(e.g., a carrier capable of suspending or dissolving the activecompound) and having the properties of being nontoxic andnon-inflammatory in a patient. Excipients may include, for example:antiadherents, antioxidants, binders, coatings, compression aids,disintegrants, dyes (colors), emollients, emulsifiers, fillers(diluents), film formers or coatings, flavors, fragrances, glidants(flow enhancers), lubricants, preservatives, printing inks, sorbents,suspending or dispersing agents, sweeteners, or waters of hydration.Exemplary excipients include, but are not limited to: butylatedhydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic),calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone,citric acid, crospovidone, cysteine, ethylcellulose, gelatin,hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose,magnesium stearate, maltitol, mannitol, methionine, methylcellulose,methyl paraben, microcrystalline cellulose, polyethylene glycol,polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben,retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch(corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide,vitamin A, vitamin E, vitamin C, and xylitol.

The formulations can be mixed with auxiliary agents which do notdeleteriously react with the active compounds. Such additives caninclude wetting agents, emulsifying and suspending agents, salt forinfluencing osmotic pressure, buffers and/or coloring substances,preserving agents, sweetening agents, or flavoring agents. Thecompositions can also be sterilized if desired.

The route of administration can be any route which effectivelytransports the active compound of the invention to the appropriate ordesired site of action, such as oral, nasal, pulmonary, buccal,subdermal, intradermal, transdermal, or parenteral, includingintravenous, subcutaneous and/or intramuscular. In one embodiment, theroute of administration is oral. In another embodiment, the route ofadministration is topical.

Dosage forms can be administered once a day, or more than once a day,such as twice or thrice daily. Alternatively, dosage forms can beadministered less frequently than daily, such as every other day, orweekly, if found to be advisable by a prescribing physician or drug'sprescribing information. Dosing regimens include, for example, dosetitration to the extent necessary or useful for the indication to betreated, thus allowing the patient's body to adapt to the treatment, tominimize or avoid unwanted side effects associated with the treatment,and/or to maximize the therapeutic effect of the present compounds.Other dosage forms include delayed or controlled-release forms. Suitabledosage regimens and/or forms include those set out, for example, in thelatest edition of the Physicians' Desk Reference, incorporated herein byreference.

Proper dosages for pediatric patients can be determined using knownmethods, including weight, age, body surface area, and models such asSimcyp® Pediatric Simulation modeling (CERTARA, Princeton, N.J.) whichcan be used to establish a pharmacokinetic approach for dosing thattakes into account patient age, ontogeny of the clearance pathways toeliminate a compound of any one of formulas (Ia) through (10, and bodysurface area (BSA). In one embodiment, the dosage form is formulated toprovide a pediatric dose from about 30% to about 100% of an adult dose,or about 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%,95%, or 100% of an adult dose.

In one embodiment, the invention provides an oral pharmaceuticalcomposition comprising structure (I) or any of the structures (Ia),(Ia′), (Ib), (Ic), (Id), (Ie), (If) or (Ig), or a pharmaceuticallyacceptable salt, isomer, hydrate, solvate or isotope thereof, togetherwith at least one pharmaceutically acceptable oral carrier, diluent, orexcipient. In another embodiment, the invention provides a topicalpharmaceutical composition comprising a compound of structure (I) or anyof the structures (Ia), (Ia′), (Ib), (Ic), (Id), (Ie), (If) or (Ig), ora pharmaceutically acceptable salt, isomer, hydrate, solvate or isotopethereof, together with at least one pharmaceutically acceptable topicalcarrier, diluent, or excipient. For example, the oral pharmaceuticalcomposition is provided to treat cholestatic pruritus, wherein thedosage regimen is, for example, once a day. In one embodiment, thetopical pharmaceutical composition is provided to treat atopicdermatitis.

In another embodiment, there are provided methods of making acomposition of a compound described herein including formulating acompound of the invention with a pharmaceutically acceptable carrier ordiluent. In some embodiments, the pharmaceutically acceptable carrier ordiluent is suitable for oral administration. In some such embodiments,the methods can further include the step of formulating the compositioninto a tablet or capsule. In other embodiments, the pharmaceuticallyacceptable carrier or diluent is suitable for parenteral administration.In some such embodiments, the methods further include the step oflyophilizing the composition to form a lyophilized preparation. In someembodiments, the composition is formulated into a pediatric dosage formsuitable for treating a pediatric subject.

In certain embodiments, the invention provides a compound havingstructure (I) or any of the structures (Ia), (Ia′), (Ib), (Ic), (Id),(Ie), (If) or (Ig), or a pharmaceutically acceptable salt, isomer,hydrate, solvate or isotope thereof. Such compounds can be synthesizedusing standard synthetic techniques known to those skilled in the art.For example, compounds of the present invention can be synthesized usingappropriately modified synthetic procedures set forth in the followingExamples and Reaction Schemes.

To this end, the reactions, processes, and synthetic methods describedherein are not limited to the specific conditions described in thefollowing experimental section, but rather are intended as a guide toone with suitable skill in this field. For example, reactions may becarried out in any suitable solvent, or other reagents to perform thetransformation[s] necessary. Generally, suitable solvents are protic oraprotic solvents which are substantially non-reactive with thereactants, the intermediates or products at the temperatures at whichthe reactions are carried out (i.e., temperatures which may range fromthe freezing to boiling temperatures). A given reaction may be carriedout in one solvent or a mixture of more than one solvent. Depending onthe particular reaction, suitable solvents for a particular work-upfollowing the reaction may be employed.

All reagents, for which the synthesis is not described in theexperimental part, are either commercially available, or are knowncompounds or may be formed from known compounds by known methods by aperson skilled in the art. The compounds and intermediates producedaccording to the methods of the invention may require purification.Purification of organic compounds is well known to a person skilled inthe art and there may be several ways of purifying the same compound. Insome cases, no purification may be necessary. In some cases, thecompounds may be purified by crystallization. In some cases, impuritiesmay be stirred out using a suitable solvent. In some cases, thecompounds may be purified by chromatography, particularly flash columnchromatography, using purpose-made or prepacked silica gel cartridgesand eluents such as gradients of solvents such as heptane, ether, ethylacetate, acetonitrile, ethanol and the like. In some cases, thecompounds may be purified by preparative HPLC using methods asdescribed.

Purification methods as described herein may provide compounds of thepresent invention which possess a sufficiently basic or acidicfunctionality in the form of a salt, such as, in the case of a compoundof the present invention which is sufficiently basic, a trifluoroacetateor formate salt, or, in the case of a compound of the present inventionwhich is sufficiently acidic, an ammonium salt. A salt of this type caneither be transformed into its free base or free acid form,respectively, by various methods known to a person skilled in the art orbe used as salts in subsequent biological assays. It is to be understoodthat the specific form of a compound of the present invention asisolated and as described herein is not necessarily the only form inwhich said compound can be applied to a biological assay in order toquantify the specific biological activity.

Chemical names were generated using the ChemDraw naming software(Version 17.0.0.206) by PerkinElmer Informatics, Inc. In some cases,generally accepted names of commercially available reagents were used inplace of names generated by the naming software.

EXAMPLES General Methods

¹H NMR (400 MHz) spectra were obtained in solution of deuterochloroform(CDCl₃), deuteromethanol (CD₃OD) or dimethyl sulfoxide—D6 (DMSO). HPLCretention times, purities, and mass spectra (LCMS) were obtained usingone of the following methods:

Method 1: Agilent 1260 Infinity II System equipped with an AgilentPoroshell 120 EC-18, 2.7 μm, 4.6×100 mm column at 30° C., using H₂O with0.1% formic acid as the mobile phase A, and MeCN with 0.1% formic acidas the mobile phase B. An ESI detector in positive mode was used. Thegradient was 5-95% mobile phase B over 12 min then held at 95% for 1.8min, then return to 10% mobile phase B over 0.2 min. The flow rate was 1mL/min.

Method 2: Agilent 1290 Infinity II System equipped with an AgilentPoroshell 120 EC-18, 1.9 μm, 2.1×50 mm column at 35° C., using H₂O with0.1% formic acid as the mobile phase A, and MeCN with 0.1% formic acidas the mobile phase B. An ESI detector in positive mode was used. Thegradient was 20-95% mobile phase B over 0.8 min then held at 95% for 0.7mins, then return to 20% mobile phase B over 0.7 min. The flow rate was0.7 mL/min.

Method 3: Agilent 1260 Infinity II System equipped with an AgilentPoroshell 120 EC-18, 2.7 μm, 4.6×100 mm column at 30° C., using H₂O with0.1% formic acid as the mobile phase A, and MeCN with 0.1% formic acidas the mobile phase B. An ESI detector in positive mode was used. Thegradient was 5-95% mobile phase B over 5 min then held at 95% for 1.8min, then return to 20% mobile phase B over 0.2 min. The flow rate was 1mL/min.

Method 4: Agilent 1200 System equipped with a Kinetex C18 50×2.1 mm (5um particles), using H₂O with 0.037% Trifluoroacetic Acid as the mobilephase A, and MeCN with 0.018% Trifluoroacetic Acid as the mobile phaseB. An ESI detector in positive mode was used. The gradient was 5% B at0.00 min and 5-90% B at 0.00-0.80 min, 90-95% B at 0.80-0.12 min, andthen 95-5% B in 0.01 min, hold on 5% B for 0.34 min, the flow rate was1.5 ml/min.

Method 5: Shimadzu SCL-10A system equipped with Agilent Eclipse XDB-C18,5 uM, 4.6×150 mm column and PE Sciex API 150 EX, using water with 0.1%trifluoroacetic acid as the mobile phase A, and methanol with 0.1%trifluoroacetic acid as the mobile phase B. The gradient was 5-95%mobile phase B over 12 min then held at 95% mobile phase B for 3 min,then return to 5% mobile phase B for 1 min. The flow rate was 1 mL/min.

Method 6: Shimadzu LCMS-2020 System equipped with an Kinetex EVO C182.1×30 mm, (5 um particles), using H₂O with 0.0375% Trifluoroacetic Acidas the mobile phase A, and MeCN with 0.01875% Trifluoroacetic Acid asthe mobile phase B. An ESI detector in positive mode was used. Thegradient was 5% B at 0.00 min and 5-95% B at 0.00-0.80 min, 95-95% B at0.80-1.2 min, and then 95-5% B in 0.01 min, hold on 5% B for 0.34 min,the flow rate was 1.5 ml/min.

The pyridine, dichloromethane (DCM), tetrahydrofuran (THF), and tolueneused in the procedures were from Aldrich Sure-Seal bottles kept undernitrogen (N₂). Other solvents were used as is. All reactions werestirred magnetically, and temperatures are external reactiontemperatures. Chromatographies were typically carried out using aCombiflash Rf flash purification system (Teledyne Isco) equipped withRedisep (Teledyne Isco) Rf Gold Normal-Phase silica gel (SiO₂) columnsor by using a similar system.

Preparative HPLC purifications were typically performed using one of thefollowing systems: 1) Waters System equipped with a Waters 2489 uv/visdetector, an Aquity QDA detector, a Waters xBridge Prep C18 5 μm OBD,30×150 mm column, and eluting with various gradients of H₂O/MeCN (0.1%formic acid) at a 30 mL/min flow rate, 2) Teledyne Isco ACCQPrep® HP150UV system equipped with a Waters xBridge Prep C18 5 μm OBD, 30×150 mmcolumn, and eluting with various gradients of H₂O/MeCN (0.1% formicacid) at a 42.5 mL/min flow rate, or 3) column: Phenomenex Synergi C18150×30 mm-4 μm; mobile phase: [H₂O (0.225% formic acid)-MeCN]; B %:55%-85%, 12 min) and were typically concentrated using a Genevac EZ-2.

The following additional abbreviations are used: ethyl acetate (EA),triethylamine (TEA), water (H2O, sodium chloride (NaCl), Hydrochloridricacid (HCl), methanol (MeOH), dimethyl sulfoxide (DMSO), silica gel(SiO₂), diisobutylaluminium hydride (DIBAL), trifluoroacetic acid (TFA),4-dimethylaminopyridine (DMAP), diphenylphosphoryl azide (DPPA), benzoylperoxide (BPO), 1,1′-bis(diphenylphosphino)ferrocene (dppf),bis(pinacolato)diboron (B₂pin₂), tetrahydrofuran (THF),1,4-diazabicyclo[2.2.2]octane bis(sulfur dioxide) adduct (DABSO),hexafluorophosphate azabenzotriazole tetramethyl uronium (HATU),hydroxybenzotriazole (HOBt), N-methyl morpholine (NMM),N-Bromosuccinimide (NBS), diisopropylethyl amine (DIPEA or DIEA),diethyl azodicarboxylate (DEAD), diisopropyl azodicarboxylate (DIAD),2-[2-(dicyclohexylphosphino)phenyl]-N-methylindole (CM-Phos), triflicacid (TfOH), kethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride(EDC), isopropanol (IPA), dimethylformamide (DMF), dimethyl acetamide(DMA), dichloromethane (DCM), 1,2-dichloroethane (DCE), acetonitrile(MeCN or ACN), 1,1′-thiocarbonyldiimidazole (TCDI), petroleum ether(PE), not determined (ND), retention time (RT), molecular weight (mw),room temperature (rt), hour (hr), and not applicable (N/A).

Example 1

Synthesis of Example 1

Step 1-1. Synthesis of tert-butyl((1S,3R)-3-(4-methoxybenzamido)cyclohexyl)carbamate

To a stirring solution of an ice-cold solution of(1S,3R)-3-Amino-1-(Boc-amino)cyclohexane (1.142 g, 1.0 eq., 5.329 mmol)in DCM (30 mL) was added with DIPEA (1.377 g, 1.9 mL, 2.0 eq., 10.66mmol), followed by slow addition of 4-methoxybenzoyl chloride (954.5 mg,758 μL, 1.05 eq., 5.595 mmol). The resulting mixture was stirred at roomtemperature. After stirring for 18 hours, the reaction mixture wasfiltered, and the filter cake was washed with DCM and dried under highvacuum to yield tert-butyl((1S,3R)-3-(4-methoxybenzamido)cyclohexyl)carbamate (1.659 g, 4.761mmol, 89.35% yield).

LCMS-ESI (m/z) calculated: 348.44; found 349.2 [M+H]⁺, RT=4.239 min(Method 3).

¹H NMR (400 MHz, DMSO-d6) δ 8.09 (d, J=7.9 Hz, 1H), 7.82 (d, J=8.4 Hz,2H), 6.97 (d, J=8.4 Hz, 2H), 6.83 (d, J=8.1 Hz, 1H), 3.80 (s, 3H),3.79-3.71 (m, 1H), 3.31-3.22 (m, 1H), 1.93 (d, J=12.0 Hz, 1H), 1.77-1.68(m, 3H), 1.38 (s, 9H), 1.31-1.17 (m, 3H), 1.11-1.01 (m, 1H).

Step 1-2. Synthesis ofN-((1R,3S)-3-aminocyclohexyl)-4-methoxybenzamide.HCl

Methoxybenzamido)cyclohexyl)carbamate (1.654 g, 1 eq., 4.747 mmol) inEtOH (40 mL) was added 1.25M hydrogen chloride in 1,4-dioxane (1.731 g,37.97 mL, 1.25 molar, 10 eq., 47.47 mmol). The resulting mixture wasstirred at 50° C. for 17 hours. The reaction mixture was directlyconcentrated to yield a white solid, which was further dried under highvacuum to yield N-((1R,3S)-3-aminocyclohexyl)-4-methoxybenzamidehydrochloride (1.331 g, 4.674 mmol, 98.46% yield), which was usedwithout further purification in the next step.

LCMS-ESI (m/z) calculated: 248.44; found 249.2 [M+H]⁺, RT=1.665 min(Method 3).

¹H NMR (400 MHz, DMSO-d6) δ 8.26 (d, J=7.9 Hz, 1H), 8.09 (s, 3H), 7.84(d, J=8.4 Hz, 2H), 6.98 (d, J=8.4 Hz, 2H), 3.91-3.81 (m, 1H), 3.80 (s,3H), 3.13-3.05 (m, 1H), 2.12 (d, J=11.6 Hz, 1H), 1.91 (d, J=12.0 Hz,1H), 1.78 (d, J=12.0 Hz, 2H), 1.44-1.21 (m, 4H).

Step 1-3. Synthesis ofN-((1R,3S)-3-((2,6-bis(trifluoromethyl)pyridin-4-yl)amino)cyclohexyl)-4-methoxybenzamide(Example 1)

To a solution of 4-chloro-2,6-bis(trifluoromethyl)pyridine (53 mg, 1eq., 0.21 mmol) and N-((1R,3S)-3-aminocyclohexyl)-4-methoxybenzamidehydrochloride (60 mg, 1 eq., 0.21 mmol) in DMSO (2 mL) was added DIPEA(0.11 g, 0.15 mL, 4 eq., 0.84 mmol). The vial was capped and theresulting solution was stirred at 130° C. After stirring for 16 hours,the reaction mixture was cooled to room temperature, filtered. Thefiltrate was purified by prep-HPLC to yieldN-((1R,3S)-3-((2,6-bis(trifluoromethyl)pyridin-4-yl)amino)cyclohexyl)-4-methoxybenzamide(46 mg, 100 μmol, 48% yield).

LCMS-ESI (m/z) calculated: 461.41; found 462.2 [M+H]⁺, RT=9.559 min(Method 1).

¹H NMR (400 MHz, DMSO-d₆) δ 8.13 (d, J=8.0 Hz, 1H), 7.82 (d, J=8.4 Hz,2H), 7.63 (d, J=8.3 Hz, 1H), 7.16 (s, 2H), 6.97 (d, J=8.4 Hz, 2H),4.03-3.93 (m, 1H), 3.80 (s, 3H), 3.77-3.68 (m, 1H), 2.10-2.03 (m, 1H),1.94-1.82 (m, 2H), 1.77 (d, J=12.3 Hz, 1H), 1.48 (q, J=13.1 Hz, 1H),1.36-1.21 (m, 2H), 1.15-1.04 (m, 1H).

Example 2 Synthesis of Example 2

Step 1-1. Synthesis of tert-butyl((1S,3R)-3-benzamidocyclohexyl)carbamate

To an ice-cold solution of (1S,3R)-3-amino-1-(Boc-amino)cyclohexane(1.238 g, 1 eq., 5.777 mmol) in DCM (30 mL) was added DIPEA (1.493 g,2.0 mL, 2 eq., 11.55 mmol), followed by addition of benzoyl chloride(852.6 mg, 704.1 μL, 1.05 eq., 6.066 mmol). The resulting mixture wasallowed to stir at room temperature for 1 hour. The reaction mixture wasfiltered, and the filter cake was washed with DCM and dried under vacuumto yield tert-butyl ((1S,3R)-3-benzamidocyclohexyl)carbamate (1.575 g,4.946 mmol, 86% yield). It was used as is in the next step.

LCMS-ESI (m/z) calculated: 318.30; found 319.2 [M+H]⁺, RT=4.237 min(Method 3).

¹H NMR (400 MHz, DMSO-d6) δ 8.26 (d, J=7.9 Hz, 1H), 7.83 (d, J=7.5 Hz,2H), 7.53-7.42 (m, 3H), 6.84 (d, J=8.0 Hz, 1H), 3.84-3.74 (m, 1H),3.31-3.23 (m, 1H), 1.95 (d, J=12.0 Hz, 1H), 1.79-1.68 (m, 3H), 1.38 (s,9H), 1.31-1.17 (m, 3H), 1.11-1.02 (m, 1H).

Step 1-2. Synthesis of N-((1R,3S)-3-aminocyclohexyl)benzamidehydrochloride

To a stirred suspension of tert-butyl((1S,3R)-3-benzamidocyclohexyl)carbamate (1.570 g, 1 eq., 4.931 mmol) in1,4-dioxane (35 mL) was added 4M hydrogen chloride in 1,4-dioxane (10eq., 49.31 mmol). The resulting mixture was stirred at 50° C. for 19hours. The reaction mixture was concentrated, dried under vacuum toyield N-((1R,3S)-3-aminocyclohexyl)benzamide hydrochloride (1.201 g,4.714 mmol, 96% yield).

LCMS-ESI (m/z) calculated: 218.30; found 219.2 [M+H]⁺, RT=0.261 min(Method 2).

¹H NMR (400 MHz, DMSO-d6) δ 8.42 (d, J=7.9 Hz, 1H), 8.11 (s, 3H), 7.85(d, J=7.5 Hz, 2H), 7.55-7.41 (m, 3H), 3.93-3.80 (m, 1H), 3.18-3.04 (m,1H), 2.14 (d, J=11.8 Hz, 1H), 1.92 (d, J=12.0 Hz, 1H), 1.79 (d, J=11.4Hz, 2H), 1.48-1.20 (m, 4H).

Step 1-3. Synthesis ofN-((1R,3S)-3-((2,6-bis(trifluoromethyl)pyridin-4-yl)amino)cyclohexyl)benzamide(Example 2)

To a solution of 4-chloro-2,6-bis(trifluoromethyl)pyridine (59 mg, 1eq., 0.24 mmol) and N-((1R,3S)-3-aminocyclohexyl)benzamide hydrochloride(60 mg, 1 eq., 0.24 mmol) in DMSO (2 mL) was added DIPEA (0.12 g, 0.16mL, 4 eq., 0.94 mmol). The vial was capped and the resulting solutionwas stirred at 130° C. After stirring for 22 hours, the reaction mixturewas cooled to room temperature, filtered. The filtrate was purified byprep-HPLC to yieldN-((1R,3S)-3-((2,6-bis(trifluoromethyl)pyridin-4-yl)amino)cyclohexyl)benzamide (2.5 mg, 5.8 μmol, 2.5% yield).

LCMS-ESI (m/z) calculated: 431.38; found 432.2 [M+H]⁺, RT=9.535 min(Method 1).

Example 3 Synthesis of Example 3

Step 1-1. Synthesis of tert-butyl((1S,4S)-4-benzamidocyclohexyl)carbamate

To an ice-cold solution of 1-N-Boc-cis-1,4-cyclohexyldiamine (500 mg, 1eq., 2.33 mmol) in DCM (6 mL) was added DIPEA (603 mg, 0.81 mL, 2 eq.,4.67 mmol), followed by slow addition of benzoyl chloride (344 mg, 284μL, 1.05 eq., 2.45 mmol). The resulting mixture was stirred at roomtemperature for 1 hour. The reaction mixture was concentrated underreduced pressure and the residue purified by flash column chromatography(0-80% EtOAc/hexanes) to yield tert-butyl((1S,4S)-4-benzamidocyclohexyl)carbamate (703 mg, 2.21 mmol, 95% yield).

LCMS-ESI (m/z) calculated: 318.19; found 319.1 [M+H]⁺, RT=4.191 min(Method 3)

¹H NMR (400 MHz, DMSO-d₆) δ 8.02 (d, J=6.8 Hz, 1H), 7.83 (d, J=7.6 Hz,2H), 7.53-7.42 (m, 3H), 6.58 (s, 1H), 3.84-3.77 (m, 1H), 3.45-3.38 (m,1H), 1.77-1.67 (m, 4H), 1.59-1.48 (m, 4H), 1.39 (s, 9H).

Step 1-2. Synthesis of N-((1s,4s)-4-aminocyclohexyl)benzamidehydrochloride

To a solution of tert-butyl ((1S,4S)-4-benzamidocyclohexyl)carbamate(699 mg, 1 eq., 2.20 mmol) in 1,4-Dioxane (5 mL) was added 4M hydrogenchloride in 1,4-dioxane (800 mg, 5.49 mL, 4.00 molar, 10 eq., 22.0mmol). The resulting mixture was stirred at room temperature for 18hours. The reaction mixture was filtered, and the filter cake was washedwith diethyl ether and dried under high vacuum to yieldN-((1S,4S)-4-aminocyclohexyl)benzamide hydrochloride (517 mg, 2.03 mmol,92% yield). It was used as is in the following step.

LCMS-ESI (m/z) calculated: 218.14; found 219.2 [M+H]⁺, RT=1.294 min(Method 3)

¹H NMR (400 MHz, DMSO-d₆) δ 8.39 (s, 1H), 8.19-7.97 (m, 4H), 7.86 (d,J=7.6 Hz, 2H), 7.55-7.42 (m, 3H), 3.93-3.84 (m, 1H), 3.17-3.06 (m, 1H),1.96-1.87 (m, 2H), 1.85-1.71 (m, 4H), 1.66-1.56 (m, 2H).

Step 1-3. Synthesis ofN-((1S,4S)-4-((2,6-bis(trifluoromethyl)pyridin-4-yl)amino)cyclohexyl)benzamide (Example 3)

To a solution of 4-chloro-2,6-bis(trifluoromethyl)pyridine (59 mg, 1eq., 0.24 mmol) and N-((1S,4S)-4-aminocyclohexyl)benzamide hydrochloride(60 mg, 1 eq., 0.24 mmol) in DMSO (2 mL) was added DIPEA (0.12 g, 0.16mL, 4 eq., 0.94 mmol). The vial was capped and the resulting solutionwas heated at 130° C. After stirring for 18 hours, the reaction mixturewas cooled to room temperature and filtered. The filtrate was purifiedby prep-HPLC to yieldN-((1S,4S)-4-((2,6-bis(trifluoromethyl)pyridin-4-yl)amino)cyclohexyl)benzamide (45.6 mg, 106 μmol, 45% yield).

LCMS-ESI (m/z) calculated: 431.00; found 432.2 [M+H]⁺, RT=9.524 min(Method 1)

¹H NMR (400 MHz, DMSO-d₆) δ 8.21 (d, J=6.5 Hz, 1H), 7.84 (d, J=7.5 Hz,2H), 7.61 (d, J=6.9 Hz, 1H), 7.54-7.42 (m, 3H), 7.19 (s, 2H), 3.94-3.84(m, 1H), 3.78-3.70 (m, 1H), 1.82-1.66 (m, 8H).

Example 4 Synthesis of Example 4

4-fluoro-2-(trifluoromethyl)benzonitrile (12 mg, 63.46 umol, 1.05 eq.),N-((1S,4S)-(4-aminocyclohexyl)-4-methoxy-benzamide (15 mg, 60.41 umol, 1eq.), K₂CO₃ (25.05 mg, 181.22 umol, 3 eq.) were dissolved in ACN (1 mL)and heated to 80° C. for 12 hours. The mixture was cooled down to roomtemperature, filtered and the filtrate was concentrated in vacuo. Theresidue was purified by prep-HPLC to yieldN-[4-[4-cyano-3-(trifluoromethyl)anilino]cyclohexyl]-4-methoxy-benzamide(2.0 mg, 4.79 umol, 8% yield).

¹H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.65 (d, J=8.93 Hz, 2H) 7.48-7.51(m, 1H) 6.85-6.88 (m, 2H) 6.78-6.80 (m, 1H) 6.61-6.64 (m, 1H) 5.88-5.94(m, 1H) 4.04-4.11 (m, 1H) 3.78-3.80 (m, 3H) 3.54-3.61 (m, 1H) 1.81-1.90(m, 4H) 1.61-1.73 (m, 4H).

The compounds listed in Table 1 were made using the procedures of Scheme1.

TABLE 1 Cpd Purity RT Observed Purity Structure No. (min) MW m/z IonMethod

1-1  0.562 449.37 450.1 [M + H]⁺ Method 4

1-2  0.978 474.45 475.3 [M + H]⁺ Method 4

1-3  0.996 461.41 462.3 [M + H]⁺ Method 4

1-4  0.994 456.39 457.1 [M + H]⁺ Method 4

1-5  1.001 435.37 436.1 [M + H]⁺ Method 4

1-6  0.854 434.46 435.3 [M + H]⁺ Method 6

1-7  9.438 412.39 413.2 [M + H]⁺ Method 1

1-8  10.283 425.43 426.2 [M + H]⁺ Method 1

1-9  10.492 451.47 452.2 [M + H]⁺ Method 1

1-10 9.932 407.44 408.3 [M + H]⁺ Method 1

1-11 0.968 436.46 437.3 [M + H]⁺ Method 6

1-12 0.894 426.44 427.1 [M + H]⁺ Method 6

1-13 1.017 448.46 449.1 [M + H]⁺ Method 6

1-14 0.935 508.56 509.2 [M + H]⁺ Method 6

1-15 0.781 409.43 410.1 [M + H]⁺ Method 6

1-16 0.889 430.47 431.2 [M + H]⁺ Method 6

1-17 0.917 391.39 392.2 [M + H]⁺ Method 6

1-18 0.937 494.53 495.3 [M + H]⁺ Method 6

1-19 0.757 426.44 427 [M + H]⁺ Method 6

1-20 0.715 427.43 428.2 [M + H]⁺ Method 6

1-21 0.966 430.47 431.2 [M + H]⁺ Method 6

1-22 0.978 405.39 406.1 [M + H]⁺ Method 6

1-23 0.981 417.43 418.1 [M + H]⁺ Method 6

1-24 0.98 444.48 445.2 [M + H]⁺ Method 6

1-25 0.965 472.51 473.3 [M + H]⁺ Method 6

Example 5 Synthesis of Example 5

Synthesis ofN-((1S,3R)-3-((3,5-bis(trifluoromethyl)phenyl)amino)cyclohexyl)-4-methoxybenzamide

A mixture of 3,5-bis(trifluoromethyl)bromobenzene (103 mg, 1 eq., 351μmop, N-((1R,3S)-3-aminocyclohexyl)-4-methoxybenzamide.HCl (100 mg, 1eq., 351 μmop, cesium carbonate (343 mg, 3 eq., 1.05 mmol),palladium(II) acetate (3.94 mg, 0.05 eq., 17.6 μmop, and BINAP (21.9 mg,0.10 eq., 35.1 μmop in 1,4-Dioxane (3 mL) was bubbled with nitrogen for5 minutes. The vial was capped and the resulting yellow mixture wasstirred at 100° C. After stirring for 18 hours, the reaction mixture wascooled to room temperature, filtered. The filtrate was purified byprep-HPLC to yieldN-((1S,3R)-3-((3,5-bis(trifluoromethyl)phenyl)amino)cyclohexyl)-4-methoxybenzamide(36.6 mg, 79.5 μmol, 22.6% yield).

LCMS-ESI (m/z) calculated: 460.42; found 461.2 [M+H]⁺, RT=10.799 min(Method 1)

¹H NMR (400 MHz, DMSO-d₆) δ 8.12 (d, J=7.9 Hz, 1H), 7.82 (d, J=8.4 Hz,2H), 7.13 (s, 2H), 7.02-6.95 (m, 3H), 6.56 (d, J=8.3 Hz, 1H), 4.00-3.90(m, 1H), 3.80 (s, 3H), 3.61-3.50 (m, 1H), 2.13-2.06 (m, 1H), 1.93 (d,J=12.5 Hz, 1H), 1.85 (d, J=12.2 Hz, 1H), 1.80-1.73 (m, 1H), 1.53-1.42(m, 1H), 1.32-1.20 (m, 2H), 1.11-1.00 (m, 1H).

Example 6 Synthesis of Example 6

Synthesis of tert-butylN-((1S,4S)-[4-[(4-fluorobenzoyl)amino]cyclohexyl]carbamate

To a mixture of tert-butyl cis-N-(4-aminocyclohexyl)carbamate (1.35 g,6.31 mmol, 1 eq.) and TEA (1.28 g, 12.61 mmol, 1.76 mL, 2 eq.) in DCM(20 mL) was added 4-fluorobenzoyl chloride (1 g, 6.31 mmol, 757.58 uL, 1eq.) at 0° C., then the reaction mixture was stirred at 25° C. for 0.5hr. The reaction mixture was concentrated to give a residue that waspurified by silica on column chromatography (eluent: PE/EtOAc=10:1 to1:1). Tert-butyl N-[4-[(4-fluorobenzoyl)amino]cyclohexyl]carbamate (2 g,5.95 mmol, 94% yield) was obtained.

Synthesis of cis-N-(4-aminocyclohexyl)-4-fluoro-benzamide.HCl

A mixture of tert-butylN-[4-[(4-fluorobenzoyl)amino]cyclohexyl]carbamate (2 g, 5.95 mmol, 1eq.) in HCl/EtOAc (4 M, 5 mL, 3.36 eq.) was stirred at 25° C. for 1 hr.The reaction mixture was concentrated to giveN-(1S,4S)-(4-aminocyclohexyl)-4-fluoro-benzamide.HCl (1.5 g, 5.50 mmol,93% yield). It was used as is in the next step directly.

LCMS-ESI (m/z) calculated: 236.3; found 237.4 [M+H]⁺, RT=0.237 min(Method 4).

Synthesis ofN-((1S,4S)-4-((3,5-bis(trifluoromethyl)phenyl)amino)cyclohexyl)-4-fluorobenzamide

A mixture of 3,5-bis(trifluoromethyl)bromobenzene (107 mg, 1 eq., 367μmop, N-(1S,4S)-(4-aminocyclohexyl)-4-fluoro-benzamide.HCl (100 mg, 1eq., 367 μmop, cesium carbonate (358 mg, 3 eq., 1.10 mmol),palladium(II) acetate (4.12 mg, 0.05 eq., 18.3 μmop, and BINAP (22.8 mg,0.10 eq., 36.7 μmop in 1,4-Dioxane (3 mL) was bubbled with nitrogen for5 minutes. The vial was capped and the resulting mixture was stirred at100° C. After stirring for 16.5 hours, the reaction mixture was cooledto room temperature, filtered. The filtrate was purified by prep-HPLC toyieldN-((1S,4S)-4-((3,5-bis(trifluoromethyl)phenyl)amino)cyclohexyl)-4-fluorobenzamide(7.8 mg, 17 μmol, 5% yield).

LCMS-ESI (m/z) calculated: 448.385; found 449.2 [M+H]⁺, RT=11.396 min(Method 1).

¹H NMR (400 MHz, DMSO-d₆) δ 8.23 (d, J=6.7 Hz, 1H), 7.93 (s, 2H), 7.28(t, J=8.8 Hz, 2H), 7.14 (s, 2H), 7.02 (s, 1H), 6.60 (d, J=6.4 Hz, 1H),3.91-3.82 (m, 1H), 3.63-3.55 (m, 1H), 1.84-1.63 (m, 8H).

Example 7 Synthesis of Example 7

Synthesis ofN-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)-4-pyridyl]amino]cyclohexyl]-4-fluoro-benzamide

A mixture of N-(1S,4S)-(4-aminocyclohexyl)-4-fluoro-benzamide.HCl(177.43 mg, 650.54 umol, 1 eq.),2-chloro-4-iodo-6-(trifluoromethyl)pyridine (200 mg, 650.54 umol, 1eq.), DavePhos (25.60 mg, 65.05 umol, 0.1 eq.) and t-BuONa (187.56 mg,1.95 mmol, 3 eq.) in dioxane (10 mL) was degassed and purged with N₂ for3 times, and then Pd₂(dba)₃ (29.79 mg, 32.53 umol, 0.05 eq.) was addedunder N₂ atmosphere. The reaction mixture was stirred at 100° C. for 3 hunder N₂ atmosphere. The reaction mixture was concentrated under reducedpressure to give a residue. The residue was purified by prep-HPLC andlyophilized to giveN-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)-4-pyridyl]amino]cyclohexyl]-4-fluoro-benzamide(19.4 mg, 46.66 umol, 7% yield).

LCMS-ESI (m/z) calculated: 415.82; found 416.2 [M+H]⁺, RT=0.974 min(Method 6).

¹H NMR (400 MHz, DMSO-d6) δ=8.22 (br d, J=6.4 Hz, 1H), 8.00-7.85 (m,2H), 7.38 (br d, J=7.0 Hz, 1H), 7.28 (t, J=9.2 Hz, 2H), 7.12 (br d,J=2.4 Hz, 1H), 7.12-6.95 (m, 1H), 6.77 (br s, 1H), 3.87 (br d, J=1.2 Hz,1H), 3.63 (br s, 1H), 1.83-1.60 (m, 8H).

Example 8 Synthesis of Example 8

Synthesis of N-(1S,4S)-[4-[[2-cyclopropyl-6-(trifluoromethyl)-4-pyridyl]amino]cyclohexyl]-4-fluoro-benzamide

A mixture of N-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)-4-pyridyl]amino]cyclohexyl]-4-fluoro-benzamide(100 mg, 240.49 umol, 1 eq.), cyclopropylboronic acid (61.97 mg, 721.48umol, 3 eq.), Cs₂CO₃ (156.71 mg, 480.99 umol, 2 eq.) in dioxane (3 mL)was degassed and purged with N2 for 3 times. Pd(dppf)Cl₂ (17.60 mg,24.05 umol, 0.1 eq.) was added under N₂ atmosphere. The mixture wasstirred at 100° C. for 12 hrs under N₂ atmosphere. The reaction mixturewas filtered, and the filtrate concentrated to give the residue. Theresidue was purified by prep-HPLC to give N-(1S,4S)-[4-[[2-cyclopropyl-6-(trifluoromethyl)-4-pyridyl]amino]cyclohexyl]-4-fluoro-benzamide(8.9 mg, 18.72 umol, 8% yield).

LCMS-ESI (m/z) calculated: 421.44; found 422.3 [M+H]⁺, RT=0.839 min(Method 4).

¹H NMR (400 MHz, DMSO-d6) δ=8.51-8.38 (m, 1H), 8.24 (d, J=6.8 Hz, 1H),7.95-7.89 (m, 2H), 7.34-7.23 (m, 2H), 6.85-6.76 (m, 2H), 6.60 (s, 1H),3.87 (br s, 1H), 3.58 (br d, J=3.2 Hz, 1H), 2.04-1.91 (m, 1H), 1.83-1.62(m, 8H), 0.92-0.79 (m, 4H).

Example 9 Synthesis of Example 9

Synthesis ofN-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)-4-pyridyl]amino]cyclohexyl]-4-methoxy-benzamide

To a solution of 2-chloro-4-iodo-6-(trifluoromethyl)pyridine (200 mg,650.54 umol, 1 eq.) andN-(1S,4S)-(4-aminocyclohexyl)-4-methoxy-benzamide (161.54 mg, 650.54umol, 1 eq.) in dioxane (5 mL) was added Pd2(dba)3 (59.57 mg, 65.05umol, 0.1 eq.), Xantphos (75.28 mg, 130.11 umol, 0.2 eq.) and Cs2CO3(423.91 mg, 1.30 mmol, 2 eq.). Then mixture was stirred at 80° C. for 12hrs under N2 atmosphere. The residue was purified by flash silica gelchromatography (petroleum ether:ethyl acetate) to yieldN-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)-4-pyridyl]amino]cyclohexyl]-4-methoxy-benzamide(90 mg, 121.86 umol, 18.73% yield).

LCMS-ESI (m/z) calculated: 427.85; found 428.2 [M+H]⁺, RT=0.937 min(Method 4).

Synthesis of4-methoxy-N-[4-[[2-(3-methylanilino)-6-(trifluoromethyl)-4-pyridyl]amino]cyclohexyl]benzamide

To a solution ofN-[4-[[2-chloro-6-(trifluoromethyl)-4-pyridyl]amino]cyclohexyl]-4-methoxy-benzamide(30 mg, 40.62 umol, 1 eq.) and 3-methylaniline (6.53 mg, 60.93 umol,6.60 uL, 1.5 eq.) in dioxane (2 mL) was added Cs₂CO₃ (26.47 mg, 81.24umol, 2 eq.) and Ruphos Pd G3 (3.40 mg, 4.06 umol, 0.1 eq.). The mixturewas stirred at 100° C. for 12 hrs under N2 atmosphere. After return tort, the mixture was filtrated, and the filtrate was concentrated undervacuum to give a residue. The residue was purified by prep-HPLC to yield4-methoxy-N-[4-[[2-(3-methylanilino)-6-(trifluoromethyl)-4-pyridyl]amino]cyclohexyl]benzamide(13.3 mg, 25.51 umol, 63% yield).

LCMS-ESI (m/z) calculated: 498.55; found 499.3 [M+H]⁺, RT=0.932 min(Method 4).

The compounds listed in Table 2 were made using the procedures of Scheme2.

TABLE 2 Purity Cpd RT Observed Purity Reaction Structure No. (min) MWm/z Ion Method Conditions

2-1 0.832 449.52 450.4 [M + H]⁺ Method 4 RuPhosPdG3/C s₂Co₃/dioxane

2-2 0.815 410.41 411 [M + H]⁺ Method 6 tBuXPhosPdG3, tBuONa, 2-Me-2-butanol

2-3 0.489 441.46 442.4 [M + H]⁺ Method 6 Pd₂dba₃, Xantphos, Cs₂CO₃,toluene

Example 10 Synthesis of Example 10

Synthesis ofN-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]-4-fluoro-benzamide

A mixture of 2,4-dichloro-6-(trifluoromethyl)pyrimidine (0.1 g, 460.88umol, 100.00 uL, 1 eq.),N-(1S,4S)-(4-aminocyclohexyl)-4-fluoro-benzamide.HCl (125.70 mg, 460.88umol, 1 eq.) and DIEA (119.13 mg, 921.76 umol, 160.55 uL, 2 eq.) in DMF(1 mL) was stirred at 25° C. for 1 hour. The reaction mixture wasdiluted with H₂O (15 mL) and extracted with EA (25 mL×3). The combinedorganic layers were combined and washed with sat.aq NaCl (50 mL×3),dried over anhydrous Na2SO4, filtered and concentrated under reducedpressure to give a residue. The residue was purified by prep-TLC (SiO2,PE:EA=1:1) to yieldN-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]-4-fluoro-benzamide(0.15 g, 359.89 umol, 78% yield).

¹H NMR (400 MHz, DMSO-d6) δ=8.45 (d, J=6.8 Hz, 1H), 8.25 (d, J=6.4 Hz,1H), 7.94-7.90 (m, 2H), 7.31-7.26 (m, 2H), 6.97 (s, 1H), 3.98-3.79 (m,2H), 1.84-1.80 (m, 2H), 1.78-1.71 (m, 6H).

Synthesis ofN-(1S,4S)-[4-[[2-anilino-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]-4-fluoro-benzamide

Aniline (11.17 mg, 119.96 umol, 10.95 uL, 1 eq.),N-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]-4-fluoro-benzamide(50 mg, 119.96 umol, 1 eq.) and DIEA (77.52 mg, 599.80 umol, 104.47 uL,5 eq.) were taken up into a microwave tube in NMP (2 mL). The sealedtube was heated at 180° C. for 8 hrs in a microwave. The reactionmixture was filtered. The filtrate was purified by prep-HPLC to yieldN-[4-[[2-anilino-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]-4-fluoro-benzamide(5 mg, 9.62 umol, 8% yield).

LCMS-ESI (m/z) calculated: 473.48; found 474.2 [M+H]⁺, RT=1.021 min(Method 4).

¹H NMR (400 MHz, DMSO-d6) δ=9.47 (s, 1H), 8.44 (s, 1H), 8.27 (d, J=6.0Hz, 1H), 7.95-7.91 (m, 2H), 7.77-7.71 (m, 3H), 7.31-7.23 (m, 4H), 6.94(t, J=7.6 Hz, 1H), 6.44 (s, 1H), 4.04 (d, J=1.2 Hz, 1H), 3.90 (d, J=1.2Hz, 1H), 1.90-1.84 (m, 2H), 1.80-1.69 (m, 6H).

Example 11 Synthesis of Example 11

Synthesis4-(dimethylamino)-N-(1S,4S)-[4-[[2-[ethyl(methyl)amino]-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]benzamide

To the mixture ofN-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]-4-(dimethylamino)benzamide.HCl(50 mg, 113.15 umol, 1 eq.), N-methylethanamine (21.63 mg, 226.31 umol,31.44 uL, 2 eq., HCl) in NMP (1 mL) was added DIEA (73.12 mg, 565.77umol, 98.55 uL, 5 eq.) at 25° C., then the reaction mixture was stirredat 120° C. for 1 hr. The reaction mixture was filtered. The filtrate waspurified by prep-HPLC to yield compound4-(dimethylamino)-N-(1S,4S)-[4-[[2-[ethyl(methyl)amino]-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]benzamide (21 mg, 45.21 umol, 40% yield).

LCMS-ESI (m/z) calculated: 464.54; found 465 [M+H]⁺, RT=0.968 min(Method 4).

¹H NMR (400 MHz, DMSO-d6) δ=7.80-7.76 (m, 1H), 7.73 (d, J=8.8 Hz, 2H),7.36 (br d, J=5.2 Hz, 1H), 6.68 (d, J=9.0 Hz, 2H), 6.21 (s, 1H), 3.97(br d, J=1.2 Hz, 1H), 3.82 (br t, J=8.4 Hz, 1H), 3.56 (q, J=7.0 Hz, 2H),3.02 (s, 3H), 2.95 (s, 6H), 1.88 (br s, 2H), 1.75-1.58 (m, 6H), 1.07 (t,J=7.0 Hz, 3H).

Example 12 Synthesis of Example 12

SynthesisN-(1S,4S)-[4-[[2-(ethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]-4-methoxy-benzamide

A mixture ofN-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]-4-methoxy-benzamide(30 mg, 69.96 umol, 1 eq.), ethanamine (6.31 mg, 139.92 umol, 9.16 uL, 2eq., HCl) and K2CO3 (48.34 mg, 349.80 umol, 5 eq.) in DMF (1 mL) wasstirred at 120° C. for 12 hrs. The reaction mixture was filtered, andthe filtrate concentrated to give a residue. The residue was purified byprep-HPLC to giveN-(1S,4S)-[4-[[2-(ethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]-4-methoxy-benzamide(3 mg, 6.49 umol, 9% yield).

LCMS-ESI (m/z) calculated: 437.47; found 438 [M+H]⁺, RT=0.868 min(Method 6).

1H NMR (400 MHz, CHLOROFORM-d) δ ppm 7.73 (d, J=8.88 Hz, 2H) 6.94 (d,J=8.75 Hz, 2H) 5.99 (s, 2H) 5.03-5.13 (m, 1H) 4.09-4.21 (m, 1H)3.90-4.04 (m, 1H) 3.86 (s, 3H) 3.36-3.47 (m, 2H) 1.84-1.97 (m, 5H)1.63-1.75 (m, 3H) 1.21 (t, J=7.25 Hz, 3H).

The compounds listed in Table 3 were made using the procedures used forExample 12 in the presence of the appropriate amine.

TABLE 3 Cpd Purity RT Observed Purity Structure No. (min) MW m/z IonMethod

3-1 0.963 465.52 466.2 [M + H]⁺ Method 6

Example 13 Synthesis of Example 13

Synthesis of tert-butyl((1S,4S)-4-((3,5-bis(trifluoromethyl)phenyl)amino) cyclohexyl)carbamate

A solution of 3,5-bis(trifluoromethyl)fluorobenzene (500 mg, 1 eq., 2.15mmol) and tert-butyl ((1S,4S)-4-aminocyclohexyl)carbamate (462 mg, 1eq., 2.15 mmol) and DIPEA (1.11 g, 1.5 mL, 4 eq., 8.62 mmol) in DMSO (8mL) was stirred at 130° C. After 24 hours, the reaction mixture wascooled to room temperature, diluted with water and extracted with EtOAc.The organic layers were combined, washed with brine and concentratedunder reduced pressure. The residue was dissolved in DCM and purified byflash column chromatography (40 g SiO₂, 0-80% EtOAc/hexanes) to yieldtert-butyl((1S,4S)-4-((3,5-bis(trifluoromethyl)phenyl)amino)cyclohexyl)carbamate(333 mg, 781 μmol, 36%).

LCMS-ESI (m/z) calculated: 426.40; found 427.1 [M+H]⁺, RT=6.335 min(Method 3).

¹H NMR (400 MHz, DMSO-d₆) δ 7.11 (s, 2H), 6.99 (s, 1H), 6.86-6.69 (m,1H), 6.52 (d, J=6.8 Hz, 1H), 3.52-3.44 (m, 1H), 3.41-3.33 (m, 1H),1.66-1.53 (m, 8H), 1.38 (s, 9H).

Synthesis of(1S,4S)-N1-(2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,4-diaminehydrochloride

To a solution containing tert-butyl((1S,4S)-4-((3,5-bis(trifluoromethyl)phenyl) amino)cyclohexyl)carbamate(329 mg, 1 eq., 804 μmop in 1,4-Dioxane (5 mL) was added 4M hydrogenchloride in 1,4-dioxane (293 mg, 2.01 mL, 4.00 molar, 10 eq., 8.04mmol). The resulting mixture was allowed to stir at room temperature for21 hours. The reaction mixture was filtered, and the filter cake waswashed with diethyl ether and dried under high vacuum to yield(1S,4S)-N1-(2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,4-diaminehydrochloride (262 mg, 758 μmol, 94% yield). It was used as is in thenext step.

LCMS-ESI (m/z) calculated: 326.79; found 327.2 [M+H]⁺, RT=3.236 min(Method 3).

¹H NMR (400 MHz, DMSO-d₆) δ 8.19-7.97 (m, 3H), 7.15 (s, 2H), 7.03 (s,1H), 6.50 (s, 3H), 3.62-3.55 (m, 1H), 3.18-3.07 (m, 1H), 1.81-1.61 (m,8H).

Synthesis ofN-((1S,4S)-4-((3,5-bis(trifluoromethyl)phenyl)amino)cyclohexyl)benzamide

To a stirred solution of(1S,4S)-N1-(2-(trifluoromethyl)quinolin-4-yl)cyclohexane-1,4-diaminehydrochloride (75 mg, 1 eq., 0.21 mmol) and DIPEA (0.11 g, 0.14 mL, 4eq., 0.83 mmol) in DMF (2 mL) was added benzoyl chloride (31 mg, 25 μL,1.05 eq., 0.22 mmol). The resulting mixture was stirred at roomtemperature for 1 hour. The reaction mixture was filtered and thefiltrate purified by prep-HPLC to yieldN-((1S,4S)-4-((3,5-bis(trifluoromethyl)phenyl)amino)cyclohexyl)benzamide (21.4 mg, 49.7 μmol, 24% yield).

LCMS-ESI (m/z) calculated: 430.39; found 431.2 [M+H]⁺, RT=11.032 min(Method 1).

¹H NMR (400 MHz, DMSO-d₆) δ 8.20 (d, J=6.7 Hz, 1H), 7.84 (d, J=7.5 Hz,2H), 7.54-7.42 (m, 3H), 7.15 (s, 2H), 7.02 (s, 1H), 6.59 (d, J=6.4 Hz,1H), 3.93-3.83 (m, 1H), 3.63-3.56 (m, 1H), 1.81-1.64 (m, 8H).

Example 14 Synthesis of Example 14

Synthesis ofN-((1S,4S)-4-((3,5-bis(trifluoromethyl)phenyl)amino)cyclohexyl)-4-methoxybenzamide

To a solution of N-((1S,4S)-4-((3,5-bis(trifluoromethyl)phenyl)amino)cyclohexyl)benzamide (75 mg, 1 eq., 0.21 mmol) and DIPEA (0.11 g, 0.14mL, 4 eq., 0.83 mmol) in DMF (2 mL) was added 4-methoxybenzoyl chloride(37 mg, 29 μL, 1.05 eq., 0.22 mmol). The resulting mixture was stirredat room temperature for 1 hour. The reaction mixture was filtered andthe filtrate purified by prep-HPLC to yieldN-((1S,4S)-4-((3,5-bis(trifluoromethyl)phenyl)amino)cyclohexyl)-4-methoxybenzamide(24.7 mg, 53.6 μmol, 26% yield).

LCMS-ESI (m/z) calculated: 460.42; found 461.2 [M+H]⁺, RT=11.039 min(Method 1).

¹H NMR (400 MHz, DMSO-d₆) δ 8.03 (d, J=6.7 Hz, 1H), 7.84 (d, J=8.5 Hz,2H), 7.14 (s, 2H), 7.03-6.94 (m, 3H), 6.59 (d, J=6.4 Hz, 1H), 3.89-3.82(m, 1H), 3.80 (s, 3H), 3.62-3.56 (m, 1H), 1.82-1.63 (m, 8H).

Example 15 Synthesis of Example 15

Synthesis ofN-(1S,4S)-[4-[[2,6-bis(trifluoromethyl)-4-pyridyl]amino]cyclohexyl]-3-(dimethylamino)benzamide

To a solution ofN-4-[2,6-bis(trifluoromethyl)-4-pyridyl]cyclohexane-1,4-diamine (50 mg,113.30 umol, 1 eq., TFA) and 3-(dimethylamino)benzoic acid (18.72 mg,113.30 umol, 1 eq.) in DMF (1.5 mL) was added DIPEA (43.93 mg, 339.91umol, 59.21 uL, 3 eq.). Then HATU (64.62 mg, 169.96 umol, 1.5 eq.) wasadded and the mixture was stirred for 2 hrs at 25° C. The mixture waspartitioned between EtOAc (25 mL) and water (25 mL). The aqueous phasewas extracted with EtOAc (10 mL×2). The combined organic layers weredried over MgSO4 and concentrated under vacuum. The crude product waspurified by reversed-phase HPLC to yieldN-(1S,4S)-[4-[[2,6-bis(trifluoromethyl)-4-pyridyl]amino]cyclohexyl]-3-(dimethylamino)benzamide (45.37 mg, 95.63 umol, 84%yield).

LCMS-ESI (m/z) calculated: 474.4; found 475.1 [M+H]⁺, RT=0.589 min(Method 4).

¹H NMR (400 MHz, DMSO-d6) δ=8.10-8.04 (m, 1H), 7.64-7.58 (m, 1H),7.29-7.18 (m, 2H), 7.14 (br s, 3H), 6.89-6.84 (m, 1H), 3.91-3.83 (m,1H), 3.78-3.72 (m, 1H), 2.94 (s, 6H), 1.73 (br s, 8H).

Example 16 Synthesis of Example 16

Synthesis of tert-butylN-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]carbamate

To the mixture of 2,4-dichloro-6-(trifluoromethyl)pyrimidine (1 g, 4.61mmol, 1.00 mL, 1 eq.), tert-butyl N-(1S,4S)-(4-aminocyclohexyl)carbamate(987.68 mg, 4.61 mmol, 1 eq.) in DMF (20 mL) was added DIEA (1.19 g,9.22 mmol, 1.61 mL, 2 eq.) at 25° C., then the reaction mixture wasstirred at 25° C. for 12 hrs. The reaction mixture was partitionedbetween ethyl acetate (30 mL) and water (20 mL), extracted with ethylacetate (30 mL×3). The combined organic layers were washed with brine(20 mL×3), dried over Na₂SO4, filtered and concentrated under reducedpressure to give a residue. The residue was purified by columnchromatography (SiO2, Petroleum ether/Ethyl acetate) to yield tert-butylN-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]carbamate(1.62 g, 3.77 mmol, 82% yield).

¹H NMR (400 MHz, DMSO-d6) δ=8.38 (br d, J=7.2 Hz, 1H), 6.95 (s, 1H),6.84 (br dd, J=1.0, 2.6 Hz, 1H), 3.96-3.83 (m, 1H), 3.38 (br d, J=4.0Hz, 1H), 1.72-1.53 (m, 8H), 1.39 (s, 9H).

Synthesis of tert-butylN-(1S,4S)-[4-[[2-(dimethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]carbamate

To the mixture of tert-butylN-(1S,4S)-[4-[[2-chloro-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]carbamate(1 g, 2.53 mmol, 1 eq.), N-methylmethanamine (413.07 mg, 5.07 mmol,464.13 uL, 2 eq., HCl) in DMSO (10 mL) was added DIEA (654.70 mg, 5.07mmol, 882.34 uL, 2 eq.) at 25° C., then the reaction mixture was stirredat 120° C. for 1 hr. The reaction mixture was diluted with water (10 mL)and extracted with ethyl acetate (20 mL×3). The combined organic layerswere washed with brine (20 mL), dried over Na₂SO4, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO2, Petroleum ether/Ethyl acetate)to yield tert-butylN-(1S,4S)-[4-[[2-(dimethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]carbamate(1 g, 2.45 mmol, 97% yield).

¹H NMR (400 MHz, DMSO-d6) δ=7.32 (br d, J=4.0 Hz, 1H), 6.83-6.70 (m,1H), 6.18 (s, 1H), 3.90 (br d, J=2.0 Hz, 1H), 3.34-3.33 (m, 1H), 3.04(s, 6H), 1.76-1.68 (m, 2H), 1.61 (br dd, J=3.8, 6.2 Hz, 2H), 1.58-1.54(m, 4H), 1.38 (s, 9H).

Synthesis ofN-(1S,4S)-4-(4-aminocyclohexyl)-N2,N2-dimethyl-6-(trifluoromethyl)pyrimidine-2,4-diamine

Tert-butylN-(1S,4S)-[4-[[2-(dimethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]carbamate (500 mg, 1.24 mmol, 1 eq.) in 4N HCl/EtOAc (10 mL) was stirredat 25° C. for 1 hr. The reaction mixture was concentrated under reducedpressure to give a residue. The residue was diluted with MeOH (20 mL),and Amberlyst A-21, ion exchange resin was added and stirred at 25° C.for 0.5 hr til the pH=7. The compound was used in the next step withoutfurther purification.

LCMS-ESI (m/z) calculated: 303.33; found 303.09 [M+H]⁺, RT=0.723 min(Method 4).

Synthesis ofN-(1S,4S)-[4-[[2-(dimethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]-4-methoxy-benzamide(Example 16)

To the mixture ofN-(1S,4S)-4-(4-aminocyclohexyl)-N2,N2-dimethyl-6-(trifluoromethyl)pyrimidine-2,4-diamine(50 mg, 164.84 umol, 1 eq.), 4-methoxybenzoic acid (25.08 mg, 164.84umol, 22.68 uL, 1 eq.) in DMF (1 mL) was added DIEA (42.61 mg, 329.68umol, 57.42 uL, 2 eq.), HATU (68.94 mg, 181.32 umol, 1.1 eq.) at 25° C.,then the reaction mixture was stirred at 25° C. for 1 hr. The reactionmixture was filtered. The filtrate was purified by prep-HPLC andlyophilized to yieldN-(1S,4S)-[4-[[2-(dimethylamino)-6-(trifluoromethyl)pyrimidin-4-yl]amino]cyclohexyl]-4-methoxy-benzamide(33.4 mg, 76.35 umol, 46% yield).

LCMS-ESI (m/z) calculated: 437.47; found 438.0 [M+H]⁺, RT=0.941 min(Method 1).

¹H NMR (400 MHz, DMSO-d6) δ=8.02 (br d, J=5.9 Hz, 1H), 7.84 (d, J=8.8Hz, 2H), 7.37 (br d, J=4.8 Hz, 1H), 6.97 (d, J=8.8 Hz, 2H), 6.21 (s,1H), 3.98 (br s, 1H), 3.84 (br s, 1H), 3.80 (s, 3H), 3.06 (s, 6H), 1.89(br d, J=4.3 Hz, 2H), 1.75-1.62 (m, 6H).

Example 17 Synthesis of Example 17

Synthesis of methyl 1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylate

To a stirring solution of methyl 1H-pyrazole-4-carboxylate (200 mg, 1eq., 1.59 mmol), 2,2,2-trifluoroethyl trifluoromethanesulfonate (736.17mg, 2 eq., 3.17 mmol), Cs₂CO₃ (1.55 g, 3 eq., 4.76 mmol) in ACN (3 mL).Then it was stirred at 60° C. for 12 hours. The mixture was filtered,and filtrate was collected. The crude product was purified byreversed-phase flash and lyophilized to yield methyl1-(2,2,2-trifluoroethyl)pyrazole-4-carboxylate (114 mg, 542.79 umol,34.23% yield).

LCMS-ESI (m/z) calculated: 208.1; found 209.2 [M+H]⁺, RT=0.392 min(Method 4).

1H NMR (400 MHz, DMSO-d6) δ=8.47 (s, 1H), 8.00 (s, 1H), 5.21 (q, J=9.2Hz, 2H), 3.76 (s, 3H).

Synthesis of 1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylic acid

To a stirring solution of methyl1-(2,2,2-trifluoroethyl)pyrazole-4-carboxylate (114 mg, 1 eq., 547.71umol) and LiOH.H₂O (68.95 mg, 3 eq., 1.64 mmol) in MeOH (1.5 mL) and H₂O(1.5 mL). The solution was stirred at 50° C. for 1 hour. The mixture wasadjusted to pH 7 with 1 M HCl and concentrated in vacuum to yield1-(2,2,2-trifluoroethyl)pyrazole-4-carboxylic acid (106 mg, crude). Thecrude product was used for next step without purification.

LCMS-ESI (m/z) calculated: 194.1; found 195.2 [M+H]⁺, RT=0.322 min(Method 4).

Synthesis of tert-butyl((1s,4s)-4-((4-cyano-3-(trifluoromethyl)phenyl)amino)cyclohexyl)carbamate

To a stirring solution of 4-fluoro-2-(trifluoromethyl)benzonitrile (10g, 1 eq., 52.88 mmol) and tert-butyl N-(4-aminocyclohexyl)carbamate(11.33 g, 1 eq., 52.88 mmol) in DMF (100 mL) was added K₂CO₃ (14.62 g, 2eq., 105.76 mmol). The mixture was stirred at 80° C. for 1 hour. Thereaction mixture was filtered and concentrated, then the mixture waspoured into Brine (150 mL) and EtOAc (150 mL). The water layer waswashed with EtOAc (100 mL×5), the organic phase was dried over Na₂SO₄,filtered and concentrated. The residue was purified by columnchromatography to yield tert-butylN-[4-[4-cyano-3-(trifluoromethyl)anilino]cyclohexyl]carbamate (13.8 g,35.99 mmol, 68% yield).

LCMS-ESI (m/z) calculated: 383.4; found 384.2 [M+H]⁺, RT=0.679 min(Method 4).

1H NMR (400 MHz, DMSO-d6) δ=7.69 (d, J=8.6 Hz, 1H), 7.08 (br d, J=7.3Hz, 2H), 6.85 (dd, J=1.8, 8.8 Hz, 1H), 6.79 (br s, 1H), 3.49 (br s, 1H),3.39 (br d, J=2.6 Hz, 1H), 1.74-1.49 (m, 9H), 1.39 (s, 10H).

4-(((1s,4s)-4-aminocyclohexyl)amino)-2-(trifluoromethyl)benzonitrile

To a stirring solution of tert-butylN-[4-[4-cyano-3-(trifluoromethyl)anilino]cyclohexyl] carbamate (5.5 g, 1eq., 14.35 mmol) in DCM (48 mL) and TFA (12 mL). The mixture was stirredat 25° C. for 12 hours. The mixture was concentrated and was washed withMTBE (20 ml×5) to yield4-[(4-aminocyclohexyl)amino]-2-(trifluoromethyl)benzonitrile (2.6 g,9.18 mmol, 64% yield).

LCMS-ESI (m/z) calculated: 283.2; found 284.0 [M+H]⁺, RT=0.457 min(Method 4).

Synthesis ofN-((1s,4s)-4-((4-cyano-3-(trifluoromethyl)phenyl)amino)cyclohexyl)-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxamide(Example 17)

To a stirring solution of 5-(trifluoromethyl)-1H-pyrazole-4-carboxylicacid (31.73 mg, 1 eq., 176.18 umol),4-[(4-aminocyclohexyl)amino]-2-(trifluoromethyl)benzonitrile (70 mg, 1eq., 176.18 umol) in DMF (1.5 mL) was added DIPEA (68.31 mg, 3 eq.,528.54 umol) and HATU (100.48 mg, 1.5 eq., 264.27 umol) and stirred for2 hours at 25° C. The reaction mixture concentrated under reducedpressure to give a residue. The crude product was purified byreversed-phase HPLC:Column and lyophilized to yieldN-[4-[4-cyano-3-(trifluoromethyl)anilino]cyclohexyl]-5-(trifluoromethyl)-1H-pyrazole-4-carboxamide(37 mg, 82.58 umol, 47% yield).

LCMS-ESI (m/z) calculated: 445.3; found 446.1 [M+H]⁺, RT=0.672 min(Method 4).

1H NMR (400 MHz, DMSO-d6) δ=14.20-12.99 (s, 1H), 8.44-8.33 (d, 1H), 7.94(d, J=6.8 Hz, 1H), 7.71 (d, J=8.8 Hz, 1H), 7.18-7.06 (m, 2H), 6.87 (dd,J=1.8, 8.8 Hz, 1H), 3.81 (br d, J=4.6 Hz, 1H), 3.57 (br s, 1H),1.81-1.56 (m, 8H).

The compounds listed in Table 4 were made using the procedures of Scheme3.

TABLE 4 Cpd Purity RT Observed Purity Structure No. (min) MW m/z IonMethod

4-1  0.679 461.461 462.2 [M + H]⁺ Method 6

4-2  0.653 420.36 421.2 [M + H]⁺ Method 6

4-3  0.6 503.45 504.2 [M + H]⁺ Method 6

4-4  0.7 460.42 461.1 [M + H]⁺ Method 6

4-5  0.603 488.43 489.2 [M + H]⁺ Method 6

4-6  0.636 470.42 471.2 [M + H]⁺ Method 6

4-7  0.645 500.4 501 [M + H]⁺ Method 6

4-8  0.562 485.43 486.2 [M + H]⁺ Method 6

4-9  0.657 488.45 489.2 [M + H]⁺ Method 6

4-10 0.612 524.48 525.2 [M + H]⁺ Method 6

4-11 0.658 484.45 485.2 [M + H]⁺ Method 6

4-12 0.726 449.401 450.1 [M + H]⁺ Method 6

4-13 0.649 456.392 457.1 [M + H]⁺ Method 6

4-14 0.691 465.82 466.2 [M + H]⁺ Method 6

4-15 0.658 500.489 501.3 [M + H]⁺ Method 6

4-16 0.654 516.49 517.2 [M + H]⁺ Method 6

4-17 0.933 450.51 451.0 [M + H]⁺ Method 6

4-18 0.947 432.45 433 [M + H]⁺ Method 6

4-19 11.57 500.489 501.4 [M + H]⁺ Method 5

4-20 10.54 488.438 489.4 [M + H]⁺ Method 5

4-21 12.77 500.489 501.4 [M + H]⁺ Method 5

4-22 12.07 470.419 471.5 [M + H]⁺ Method 5

4-23 11.52 538.51 539.4 [M + H]⁺ Method 5

4-24 12.56 525.499 526.4 [M + H]⁺ Method 5

4-25 10.54 497.445 498.5 [M + H]⁺ Method 5

4-26 10.97 500.489 501.3 [M + H]⁺ Method 5

4-27 11.2 453.41 454.3 [M + H]⁺ Method 5

4-28 11.46 447.381 448.3 [M + H]⁺ Method 5

4-29 11.2 398.353 399.3 [M + H]⁺ Method 5

4-30 11.71 499.421 500.1 [M + H]⁺ Method 5

4-31 12.12 497.445 498.5 [M + H]⁺ Method 5

4-32 12.64 500.27 501.2 [M + H]⁺ Method 5

4-33 11.71 527.471 528.3 [M + H]⁺ Method 5

4-34 10.45 500.489 501.5 [M + H]⁺ Method 5

4-35 12.4 471.407 472.4 [M + H]⁺ Method 5

4-36 9.96 453.433 454.4 [M + H]⁺ Method 5

4-37 12.79 474.451 475.5 [M + H]⁺ Method 5

4-38 11.44 564.55 565.5 [M + H]⁺ Method 5

4-39 10.45 471.407 472.2 [M + H]⁺ Method 5

4-40 9.93 438.418 439.3 [M + H]⁺ Method 5

4-41 12.98 500.489 501.3 [M + H]⁺ Method 5

4-42 13.21 485.474 486.4 [M + H]⁺ Method 5

4-43 0.962 443.418 444.1 [M + H]⁺ Method 6

4-44 0.672 445.369 446.4 [M + H]⁺ Method 6

4-45 0.582 411.81 412.2 [M + H]⁺ Method 6

4-46 0.583 417.436 418.3 [M + H]⁺ Method 6

4-47 0.994 416.448 417.3 [M + H]⁺ Method 6

4-48 0.962 444.458 445.3 [M + H]⁺ Method 6

4-49 0.991 427.431 428 [M + H]⁺ Method 6

4-50 0.971 427.431 428.3 [M + H]⁺ Method 6

4-51 1.003 467.496 468.3 [M + H]⁺ Method 6

4-52 0.892 427.431 428 [M + H]⁺ Method 6

4-53 1.034 466.456 467.3 [M + H]⁺ Method 6

4-54 0.992 480.483 481.2 [M + H]⁺ Method 6

4-55 1.021 445.49 446.4 [M + H]⁺ Method 6

4-56 0.572 402.381 403.3 [M + H]⁺ Method 6

4-57 0.554 427.431 428.1 [M + H]⁺ Method 6

4-58 0.865 428.419 429 [M + H]⁺ Method 6

4-59 0.945 428.419 429.3 [M + H]⁺ Method 6

4-60 0.863 441.458 442 [M + H]⁺ Method 6

4-61 0.91 441.458 442 [M + H]⁺ Method 6

4-62 0.635 440.47 441.1 [M + H]⁺ Method 6

4-63 0.513 427.431 428.1 [M + H]⁺ Method 6

4-64 0.456 427.431 428.2 [M + H]⁺ Method 6

4-65 0.612 444.458 445.2 [M + H]⁺ Method 6

4-66 0.459 417.436 418.2 [M + H]⁺ Method 6

Example 18

Synthesis of Example 18 Synthesis of4-bromo-2-methyl-6-(trifluoromethyl)benzonitrile

To a stirring solution of 4-bromo-1-fluoro-2-(trifluoromethyl)benzene(10.0 g, 1 eq., 41.15 mmol) in THF (100 mL) was added LDA (24.69 mL, 1.2eq., 2 M) dropwise at −78° C. under N₂ atmosphere. After stirring at−78° C. for 0.5 hours, Mel (11.68 g, 2 eq., 82.31 mmol) was addeddropwise and the reaction was stirred at −78° C. for 0.5 hour. Thereaction solution was quenched by saturated NH₄Cl aqueous solution (200mL) and then extracted with ethyl acetate (3×100 mL). The combinedorganic layers were washed with brine (100 mL), dried over anhydroussodium sulfate, filtered and concentrated under vacuum. The residue waspurified by column chromatography and the fractions were checked by TLCto yield 5-bromo-2-fluoro-1-methyl-3-(trifluoromethyl)benzene (8.4 g,32.68 mmol, 72% yield) as colorless oil.

¹H NMR (400 MHz, CHLOROFORM-d) δ=7.46 (m, 2H), 2.24 (s, 3H).

Synthesis of 4-bromo-2-methyl-6-(trifluoromethyl)benzonitrile

To a stirring solution of5-bromo-2-fluoro-1-methyl-3-(trifluoromethyl)benzene (8.4 g, 1.0 eq.,32.68 mmol) in DMSO (100 mL) was added NaCN (1.82 g, 1.14 eq., 37.14mmol) and stirred for 2 hours at 100° C. The reaction solution wascooled to room temperature and then diluted with water (500 mL),basified to pH=10 by 1M NaOH then extracted with ethyl acetate (3×100mL). The combined organic layers were washed with brine (2×100 mL),dried over anhydrous sodium sulfate, filtered and concentrated undervacuum. The water phase was quenched by 5% NaClO aqueous solution. Theresidue was purified by column chromatography and the fractions werechecked by TLC to yield 4-bromo-2-methyl-6-(trifluoromethyl)benzonitrile(3.4 g, 12.88 mmol, 39% yield).

¹H NMR (400 MHz, CHLOROFORM-d) δ=7.77 (s, 1H), 7.74 (s, 1H), 2.64 (s,3H).

Synthesis of tert-butyl((1s,4s)-4-((4-cyano-3-methyl-5-(trifluoromethyl)phenyl)amino)cyclohexyl)carbamate

To a stirring solution of4-bromo-2-methyl-6-(trifluoromethyl)benzonitrile (1.5 g, 1 eq., 5.68mmol) tert-butyl N-(4-aminocyclohexyl)carbamate (1.22 g, 1 eq., 5.68mmol) in dioxane (20 mL) was added Cs₂CO₃ (5.55 g, 3 eq., 17.04 mmol),XantPhos Pd G3 (538.75 mg, 0.1 eq., 568.09 umol) and was stirred for 12hours at 100° C. The mixture was partitioned between EtOAc (50 mL) andwater (50 mL). The aqueous phase was extracted with EtOAc (50 mL×2).Organic phases were dried over MgSO₄ and concentrated under vacuum. Theresidue was purified by column chromatography to yield tert-butylN-[4-[4-cyano-3-methyl-5-(trifluoromethyl)anilino]cyclohexyl]carbamate(1.4 g, 3.52 mmol, 43% yield).

LCMS-ESI (m/z) calculated: 397.2; found 398.2 [M+H]⁺, RT=0.624 min(Method 4).

The compounds listed in Table 5 were made using the procedures of Scheme4 starting from tert-butyl ((1S,4S)-4-((4-cyano-3-methyl-5-(trifluoromethyl)phenyl)amino)cyclohexyl)carbamate and the appropriate acid or acid chloride.

TABLE 5 Cpd Purity RT Observed Purity Structure No. (min) MW m/z IonMethod

5-1  0.424 430.475 431.2 [M + H]+ Method 6

5-2  0.412 459.517 460.3 [M + H]+ Method 6

5-3  0.516 430.475 431.2 [M + H]+ Method 6

5-4  0.388 431.463 432.2 [M + H]+ Method 6

5-5  0.492 473.423 474.3 [M + H]+ Method 6

5-6  0.498 419.452 420.3 [M + H]+ Method 6

5-7  0.53 494.51 495.1 [M + H]+ Method 6

5-8  0.509 480.483 481.3 [M + H]+ Method 6

5-9  0.408 441.458 442.3 [M + H]+ Method 6

5-10 0.606 469.46 470 [M + H]+ Method 6

Example 19 MRGPRX2 Activity

CHO cells stably transfected to express human MRGPRX2 were maintained inan incubator at 37° C. with 5% CO₂ and grown in F12 (HAM) media with 10%fetal bovine serum (FBS), 1% Glutamax, 1% penicillin/streptomycin, 800μg/mL Geneticin (G418), and 300 μg/mL Hygromycin B. Cells were plated ina 384-well assay plate at 20,000 cells per well in 12 μL of Opti-MEM andkept in an incubator overnight. On the day of the assay, compoundssolubilized at 10 mM in DMSO were added as a 10-point curve (30 uM finaltop concentration with 1:3 serial dilutions) using a Tecan D300E digitaldispenser. Agonists were diluted in assay buffer (final concentrationsof 5.7 mM Tris-HCl, 43 mM NaCl, 50 mM LiCl, pH=8) and 2 μL of theagonist Cortistatin-14 (CPC Scientific, catalog CORT-002) are added toeach well. Final concentrations of agonists were 0.3 μM Cortistatin-14.Final concentrations of DMSO were kept consistent across the plate.Plates were incubated in the dark for 1 hr at 37° C. and then for 1 hrat room temperature. IP-1 standards and HTRF detection reagents wereadded according to the IP-One—Gq Kit purchased from Cisbio (part number62IPAPEJ) and incubated in the dark for 1 hr at room temperature. Theplate was read on a Molecular Devices SpectraMax iD5 plate reader. TheHTRF ratio was calculated from the raw data and graphed using GraphPadPrism to calculate an IC₅₀ value for each compound.

Activity data for selected MRGPRX2 antagonists (versus 0.3 uMCorstitatin-14 agonist) are displayed in Table 5. The activity rangesare denoted as follows: “+++++” denotes antagonist activity <100 nM;“++++” denotes antagonist activity between 100 and 500 nM; “+++” denotesactivity between 501 and 1000 nM; “++” denotes activity between 1001 and2500 nM; and “+” denotes activity >2500 nM.

TABLE 5 MRGPR2 Antagonist Cpd No. Activity 1-1 ++++ 1-2 ++++ 1-3 ++++1-4 ++ 1-5 ++ 1-6 ++ 1-7 ++ 1-8 +++ 1-9 ++ 1-10 +++ 1-11 ++ 1-12 +++++1-13 ++++ 1-14 +++++ 1-15 ++++ 1-16 ++++ 1-17 ++++ 1-18 ++++ 1-19 +++++1-20 ++++ 1-21 +++++ 1-22 ++++ 1-23 ++++ 1-24 ++ 1-25 ++++ 2-1 ++ 2-2+++ 2-3 +++++ 3-1 ++ 4-1 ++++ 4-2 ++++ 4-3 ++++ 4-4 ++++ 4-5 ++++ 4-6++++ 4-7 ++ 4-8 ++++ 4-9 ++++ 4-10 ++++ 4-11 ++++ 4-12 +++ 4-13 + 4-14+++ 4-15 ++ 4-16 +++ 4-17 +++ 4-18 ++ 4-19 ++ 4-20 +++ 4-21 +++ 4-22++++ 4-23 ++++ 4-24 +++ 4-25 ++++ 4-26 ++ 4-27 ++++ 4-28 ++++ 4-29 ++4-30 ++++ 4-31 ++++ 4-32 +++ 4-33 ++++ 4-34 ++ 4-35 ++ 4-36 ++ 4-37 +++4-38 ++++ 4-39 ++++ 4-40 ++ 4-41 ++++ 4-42 +++ 4-43 ++++ 4-44 +++++ 4-45+++++ 4-46 +++++ 4-47 +++++ 4-48 +++++ 4-49 +++++ 4-50 +++++ 4-51 +++++4-52 +++++ 4-53 +++++ 4-54 +++++ 4-55 +++++ 4-56 +++++ 4-57 +++++ 4-58+++++ 4-59 +++++ 4-60 +++++ 4-61 +++++ 4-62 +++++ 4-63 +++++ 4-64 +++++4-65 +++++ 4-66 +++++ 5-1 +++++ 5-2 +++++ 5-3 +++++ 5-4 +++++ 5-5 +++++5-6 +++++ 5-7 +++++ 5-8 +++++ 5-9 +++++ 5-10 +++++ example 1 + Example2 + example 3 +++ Example 4 ++++ example 5 ++ Example 6 ++ Example 7 +++Example 8 ++ Example 9 ++ Example 10 ++ Example 11 ++++ Example 12 +Example 13 + Example 14 + Example 15 ++++ Example 16 +++ Example 17+++++

The various embodiments described above can be combined to providefurther embodiments. All of the U.S. patents, U.S. patent applicationpublications, U.S. patent applications, foreign patents, foreign patentapplications and non-patent publications referred to in thisspecification and/or listed in the Application Data Sheet areincorporated herein by reference, in their entirety. Aspects of theembodiments can be modified, if necessary, to employ concepts of thevarious patents, applications and publications to provide yet furtherembodiments.

These and other changes can be made to the embodiments in light of theabove-detailed description. In general, in the following claims, theterms used should not be construed to limit the claims to the specificembodiments disclosed in the specification and the claims but should beconstrued to include all possible embodiments along with the full scopeof equivalents to which such claims are entitled. Accordingly, theclaims are not limited by the disclosure.

This application claims the benefit of priority to U.S. ProvisionalApplication No. 63/123,411, filed Dec. 9, 2020, which application ishereby incorporated by reference in its entirety.

1. A compound having structure (I):

or a pharmaceutically acceptable salt, isomer, hydrate, solvate orisotope thereof, wherein: R¹ is cycloalkyl, aryl, heterocyclyl,—(CH₂)_(n)Q, —CHQR, or —CQ(R)₂, wherein R¹ is optionally substitutedwith one or more R^(q1); Q is C₁₋₆ alkyl, aryl, cycloalkyl,heterocyclyl, —CH₂C(O)OR, —C(O)OR, —C(O)NHR, trihalomethyl,dihalomethyl, —CN, —N(R)₂, —N(R)C(O)R, —N(R)C(O)OR, or —N(R)S(O)₂R,wherein Q is optionally substituted with one or more R^(q2); R^(q1) andR^(q2) are independently H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,aryl, cycloalkyl, heteroaryl, heterocyclyl, —OR, —O(CH₂)_(n)R,haloalkoxy, —C(O)OR, —C(O)R, —OC(O)R, halo, haloalkyl, —CN, —N(R)₂,—N(R)C(═NH)N(R)₂, —N(R)C(O)R, —N(R)S(O)₂R, S(O)₂R, —C(H)Q′R, or—(CH₂)_(n)Q′ where Q′ is selected from C₁₋₆ alkyl, aryl, cycloalkyl,heterocyclyl, OR′, —C(O)OR′, —OC(O)R′, haloalkyl, —CN, —N(R′)₂,—N(R′)C(O)R′, and —N(R′)S(O)₂R′; D is N or CR^(d); W is N or CR^(w); Zis N or CR^(z); each R is independently H, C₁₋₆ alkyl, C₂₋₆ alkenyl,C₂₋₆ alkynyl, —C(O)NHR′, amino, —(CH₂)_(n)R′, halo, aryl, cycloalkyl,heteroaryl or heterocyclyl, or two R groups taken together with the atomto which they are attached form a carbocycle or heterocycle; R′ is H,C₁₋₆ alkyl, haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, cycloalkyl,heteroaryl, or heterocyclyl; R^(d) is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl, —CN, halo, —C(O)R, aryl, cycloalkyl, heteroaryl, orheterocyclyl; R^(w) is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, —CN,halo, —C(O)R, aryl, cycloalkyl, heteroaryl, or heterocyclyl; R^(z) is H,C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, —CN, halo, —C(O)R, aryl,cycloalkyl, heteroaryl, or heterocyclyl; each n is independently 0, 1,2, 3, 4 or 5; when D is CR^(d)′, W is CR^(w), and Z is CR^(z), then R²is H, C₁₋₆ alkyl, C₂₋₆ alkenyl, cycloalkyl, —OR, —C(O)OR, —OC(O)R, halo,—CF₃, —CF₂H, —C(F)H₂, C(F)₂R, —C(F)(R)₂, —N(R)₂, —N(R)C(O)R,—N(R)S(O)₂R, or S(O)₂R; or when at least one of D, W or Z is N, then R²is C₁₋₆ alkyl, C₂₋₆ alkenyl, cycloalkyl, —OR, —C(O)OR, —OC(O)R, halo,—CF₃, —CF₂H, —C(F)H₂, C(F)₂R, —C(F)(R)₂, —N(R)₂, —N(R)C(O)R,—N(R)S(O)₂R, or S(O)₂R; each R³ is independently H, C₁₋₆ alkyl, C₂₋₆alkenyl, C₂₋₆ alkynyl, aryl, cycloalkyl, heteroaryl, heterocyclyl, —OR,—C(O)OR, —OC(O)R, halo, haloalkyl, —CN, —N(R)₂, —N(R)C(O)R, —N(R)S(O)₂R,or S(O)₂R; with the provisos that: when D is CR^(d), then W and Z cannotboth be N; when D and Z are both N, then R² cannot be dimethylamino; andwhen D is CH, Z is CH and W is CR^(w), then R^(w) is not cyano orpiperidine.
 2. The compound of claim 1, wherein the compound has one ofthe following structures (Ia) or (Ia′):

or a pharmaceutically acceptable salt, isomer, hydrate, solvate orisotope thereof.
 3. The compound of claim 1, wherein the compound hasone of the following structures (Ib), (Ic), (Id), (Ie), (If) or (Ig):

or a pharmaceutically acceptable salt, isomer, hydrate, solvate orisotope thereof.
 4. The compound of claim 1, wherein R¹ is aryl,—(CH₂)_(n)Q, —CHQR, or —CQ(R)₂, wherein R¹ is optionally substitutedwith one or more R^(q1); Q is aryl, —CH₂C(O)OR, —C(O)OR, —C(O)NHR,haloalkyl, —CN, —N(R)₂, —N(R)C(O)R, —N(R)C(O)OR, or —N(R)S(O)₂R, whereinQ is optionally substituted with one or more R^(q2).
 5. The compound ofclaim 4, wherein R¹ is phenyl.
 6. The compound of claim 5, whereinphenyl is unsubstituted.
 7. The compound of claim 5, wherein phenyl issubstituted with one or more R^(q1).
 8. The compound of claim 7, whereinR^(q1) is H, —OR, —N(R)₂, —N(R)S(O)₂R, —CN, —N(R)C(O)R, —C(H)Q′R,heterocyclyl, or halo.
 9. The compound of claim 1, wherein R¹ isheterocyclyl, —(CH₂)_(n)Q, —CHQR, or —CQ(R)₂, wherein R¹ is optionallysubstituted with one or more R^(q1); Q is heterocyclyl, —CH₂C(O)OR,—C(O)OR, —C(O)NHR, haloalkyl, —CN, —N(R)₂, —N(R)C(O)R, —N(R)C(O)OR, or—N(R)S(O)₂R, wherein Q is optionally substituted with one or moreR^(q2).
 10. The compound of claim 9, wherein the heterocyclyl isunsubstituted.
 11. The compound of claim 10, wherein the heterocyclyl issubstituted with one or more R^(q1).
 12. The compound of claim 11,wherein R^(q1) is H, C₁₋₆ alkyl, —N(R)₂, or cycloalkyl.
 13. The compoundof claim 1, wherein R¹ is —(CH₂)_(n)Q; Q is —N(R)₂, —N(R)C(O)R,—N(R)C(O)OR, or —N(R)S(O)₂R, wherein Q is optionally substituted withone or more R^(q2).
 14. The compound of claim 13, wherein n is 0 and Qis N(R)₂.
 15. The compound of claim 1, wherein R¹ is —CQ(R)₂; Qis—trihalomethyl, —CN, —N(R)₂, —N(R)C(O)R, —N(R)C(O)OR, or —N(R)S(O)₂R,wherein Q is optionally substituted with one or more R^(q2).
 16. Thecompound of claim 15, wherein Q is trihalomethyl.
 17. The compound ofclaim 1, wherein each R³ is H.
 18. The compound of claim 1, wherein D isCR^(d), W is CR^(w) and Z is CR^(z) and wherein each R^(d), R^(w) andR^(z) are independently C₁₋₆ alkyl, H, —CN, halo, or —C(O)R.
 19. Thecompound of claim 1, wherein D is N, W is CR^(w) and Z is CR^(z) andwherein R² is —CF₃, C₁₋₆ alkyl, —N(R)₂, cycloalkyl, —C(O)OR, or halo.20. The compound of claim 19, wherein R² is —CF₃.
 21. The compound ofclaim 19, wherein R² is C₁₋₆ alkyl.
 22. The compound of claim 19,wherein R² is —N(R)₂.
 23. The compound of claim 19, wherein R² iscycloalkyl.
 24. The compound of claim 19, wherein R² is —C(O)OR.
 25. Thecompound of claim 19, wherein R² is halo.
 26. The compound of claim 1,wherein at least two of D, W and Z is N, and wherein R² is C₁₋₆ alkyl,—OR, or —N(R)₂.
 27. A compound selected from any one of the compoundslisted in Table 1, or a pharmaceutically acceptable salt, isomer,hydrate, solvate or isotope thereof.
 28. A pharmaceutical compositioncomprising the compound of claim 1, or a pharmaceutically acceptablesalt, isomer, hydrate, solvate or isotope thereof and at least onepharmaceutically acceptable excipient.
 29. A method of modulating aMas-Related G-Protein Receptor (MRGPR) X2 or a MRGPRX2 ortholog bycontacting MRGPRX2 or MRGPRX2 ortholog with an effective amount of thepharmaceutical composition of claim
 28. 30. A method of treating aMRGPRX2 or a MRGPRX2 ortholog dependent condition by administering to asubject in need thereof an effective amount of the pharmaceuticalcomposition of claim
 28. 31. The method of claim 30, wherein the MRGPRX2or the MRGPRX2 ortholog dependent condition is a pseudo-allergicreaction, an itch associated condition, a pain associated condition, oran inflammatory or autoimmune disorder.
 32. The method of claim 31,wherein the itch associated condition is chronic itch; contactdermatitis; Allergic blepharitis; Anemia; Atopic dermatitis; Bullouspemphigoid; Candidiasis; Chicken pox; end-stage renal failure;hemodialysis; Chronic urticaria; Contact dermatitis, Atopic Dermatitis;Dermatitis herpetiformis; Diabetes; Drug allergy, Dry skin; Dyshidroticdermatitis; Ectopic eczema; Eosinophilic fasciitis; Epidermolysisbullosa; Erythrasma; Food allergy; Folliculitis; Fungal skin infection;Hemorrhoids; Herpes; HIV infection; Hodgkin's disease; Hyperthyroidism;Iodinated contrast dye allergy; Iron deficiency anemia; Kidney disease;Leukemia, porphyrias; Lymphoma; Malignancy; Mastocystosis; Multiplemyeloma; Neurodermatitis; Onchocerciasis; Paget's disease; Pediculosis;Polycythemia rubra vera; Prurigo nodularis; Lichen Planus; LichenSclerosis; Pruritus ani; Pseudorabies; Psoriasis; Rectal prolapse;Sarcoidosis granulomas; Scabies; Schistosomiasis; Scleroderma, Severestress, Stasia dermatitis; Swimmer's itch; Thyroid disease; Tineacruris; Rosacea; Cutaneous amyloidosis; Scleroderma; Acne; woundhealing; burn healing; ocular itch; or Urticaria.
 33. The method ofclaim 32, wherein the itch associated condition is urticaria, pruritus,atopic dermatitis, dry skin, psoriasis, contact dermatitis, or eczema.34. The method of claim 31, wherein the pain associated condition isAcute Pain, Advanced Prostate Cancer, AIDS-Related Pain, AnkylosingSpondylitis, Arachnoiditis, Arthritis, Arthrofibrosis, Ataxic CerebralPalsy, Autoimmune Atrophic Gastritis, Avascular Necrosis, Back Pain,Behcet's Disease (Syndrome), Burning Mouth Syndrome, Bursitis, CancerPain, Carpal Tunnel, Cauda Equina Syndrome, Central Pain Syndrome,Cerebral Palsy, Cervical Stenosis, Charcot-Marie-Tooth (CMT) Disease,Chronic Fatigue Syndrome (CFS), Chronic Functional Abdominal Pain(CFAP), Chronic Pain, Chronic Pancreatitis, Chronic Pelvic PainSyndrome, Collapsed Lung (Pneumothorax), Complex Regional Pain Syndrome(RSD), Corneal Neuropathic Pain, Crohn's Disease, Degenerative DiscDisease, Dental Pain, Dercum's Disease, Dermatomyositis, DiabeticPeripheral Neuropathy (DPN), Dystonia, Ehlers-Danlos Syndrome (EDS),Endometriosis, Eosinophilia-Myalgia Syndrome (EMS), Erythromelalgia,Fibromyalgia, Gout, Headaches, Herniated disc, Hydrocephalus,Intercostal Neuraligia, Interstitial Cystitis, Irritable Bowel syndrome(IBS), Juvenile Dermatositis (Dermatomyositis), Knee Injury, Leg Pain,Loin Pain-Haematuria Syndrome, Lupus, Lyme Disease, Medullary SpongeKidney (MSK), Meralgia Paresthetica, Mesothelioma, Migraine,Musculoskeletal pain, Myofascial Pain, Myositis, Neck Pain, NeuropathicPain, Occipital Neuralgia, Osteoarthritis, Paget's Disease, ParsonageTurner Syndrome, Pelvic Pain, Periodontitis Pain, Peripheral Neuropathy,Phantom Limb Pain, Pinched Nerve, Polycystic Kidney Disease, PolymyalgiaRhuematica, Polymyositis, Porphyria, Post Herniorraphy Pain Syndrome,Post Mastectomy, Postoperative Pain, Pain Syndrome, Post Stroke Pain,Post Thorocotomy Pain Syndrome, Postherpetic Neuralgia (Shingles),Post-Polio Syndrome, Primary Lateral Sclerosis, Psoriatic Arthritis,Pudendal Neuralgia, Radiculopathy, Raynaud's Disease, RheumatoidArthritis (RA), Sacroiliac Joint Dysfunction, Sarcoidosi, Scheuemann'sKyphosis Disease, Sciatica, Scoliosis, Shingles (Herpes Zoster),Sjogren's Syndrome, Spasmodic Torticollis, Sphincter of OddiDysfunction, Spinal Cerebellum Ataxia (SCA Ataxia), Spinal Cord Injury,Spinal Stenosis, Syringomyelia, Tarlov Cysts, Transverse Myelitis,Trigeminal Neuralgia, Neuropathic Pain, Ulcerative Colitis, VascularPain or Vulvodynia
 35. The method of claim 31, wherein the inflammatoryor autoimmune disorder is chronic inflammation, mast cell activationsyndrome, Multiple Sclerosis, Steven Johnson's Syndrome, Toxic EpidermalNecrolysis, appendicitis, bursitis, cutaneous lupus, colitis, cystitis,dermatitis, phlebitis, reflex sympathetic dystrophy/complex regionalpain syndrome (rsd/crps), rhinitis, tendonitis, tonsillitis, acnevulgaris, sinusitis, rosacea, psoriasis, graft-versus-host disease,reactive airway disorder, asthma, airway infection, autoinflammatorydisease, celiac disease, chronic prostatitis, diverticulitis,glomerulonephritis, hidradenitis suppurativa, hypersensitivities,intestinal disorder, epithelial intestinal disorder, inflammatory boweldisease, irritable bowel syndrome, Crohn's Disease, ulcerative colitis,lupus erythematous, interstitial cystitis, otitis, pelvic inflammatorydisease, endometrial pain, reperfusion injury, rheumatic fever,rheumatoid arthritis, sarcoidosis, transplant rejection, psoriasis, lunginflammation, chronic obstructive pulmonary disease, cardiovasculardisease, or vasculitis.